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mouse anti-chicken integrin α6  (Developmental Studies Hybridoma Bank)


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    Structured Review

    Developmental Studies Hybridoma Bank mouse anti-chicken integrin α6
    <t>Integrin</t> <t>α6</t> is expressed in cortical lens, primary chick lens culture, and CEF cells. (A) Crude membrane extracts were prepared from whole chick lens, CEF cells, and differentiated primary chick lens culture containing lentoids and immunoblotted with <t>anti-integrin</t> <t>α6</t> antibody and β-actin antibody. (B) Crude membrane extracts were prepared from whole chick lens (W) or isolated E18 chick lens portions containing cortical fibers (CF) or nuclear fibers (NF). Integrin α6 was detected in the cortical lens fibers, but not in the central, nuclear lens fiber. As no actin was expressed in nuclear lens, Ponceau S staining was used to show that a comparable amount of protein was loaded on SDS-PAGE.
    Mouse Anti Chicken Integrin α6, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/chicken+%CE%B16+integrin/anti+integrin+monoclonal+antibodies/pmc07668387-230-0-11
    Average 90 stars, based on 1 article reviews
    mouse anti-chicken integrin α6 - by Bioz Stars, 2026-09
    90/100 stars

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    1) Product Images from "Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens"

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.202002154

    Integrin α6 is expressed in cortical lens, primary chick lens culture, and CEF cells. (A) Crude membrane extracts were prepared from whole chick lens, CEF cells, and differentiated primary chick lens culture containing lentoids and immunoblotted with anti-integrin α6 antibody and β-actin antibody. (B) Crude membrane extracts were prepared from whole chick lens (W) or isolated E18 chick lens portions containing cortical fibers (CF) or nuclear fibers (NF). Integrin α6 was detected in the cortical lens fibers, but not in the central, nuclear lens fiber. As no actin was expressed in nuclear lens, Ponceau S staining was used to show that a comparable amount of protein was loaded on SDS-PAGE.
    Figure Legend Snippet: Integrin α6 is expressed in cortical lens, primary chick lens culture, and CEF cells. (A) Crude membrane extracts were prepared from whole chick lens, CEF cells, and differentiated primary chick lens culture containing lentoids and immunoblotted with anti-integrin α6 antibody and β-actin antibody. (B) Crude membrane extracts were prepared from whole chick lens (W) or isolated E18 chick lens portions containing cortical fibers (CF) or nuclear fibers (NF). Integrin α6 was detected in the cortical lens fibers, but not in the central, nuclear lens fiber. As no actin was expressed in nuclear lens, Ponceau S staining was used to show that a comparable amount of protein was loaded on SDS-PAGE.

    Techniques Used: Membrane, Isolation, Staining, SDS Page

    Cx50 colocalized with integrin α6 in lens epithelium and outer cortical fiber cells. (A) Embryonic day 18 mouse lens sagittal cryosections were immunostained with anti-Cx46 (green), Cx50 (green), or integrin α6 (red) antibody and counterstained with DAPI (blue), followed by corresponding secondary antibodies conjugated with either Alexa Fluor 488 or rhodamine. The corresponding images were merged (Overlay). Scale bar, 100 µm. (B and C) Anterior epithelial cells (B) and equatorial epithelial and nascent fiber cells (C). Scale bar, 20 µm. (D) Different depths of the lens regions from equator as illustrated in panels a (equator), b (cortical), and c (nucleus), Cx50 and integrin α6 were double immunostained (left), and the extent of colocalization was quantified and presented as colocalization coefficiency using ImageJ (right). Thick black outline indicates the lens capsule. Scale bar, 20 µm. Each data point in the graph represents an individual mouse in each group. The data are presented as mean ± SEM. ***, P < 0.001.
    Figure Legend Snippet: Cx50 colocalized with integrin α6 in lens epithelium and outer cortical fiber cells. (A) Embryonic day 18 mouse lens sagittal cryosections were immunostained with anti-Cx46 (green), Cx50 (green), or integrin α6 (red) antibody and counterstained with DAPI (blue), followed by corresponding secondary antibodies conjugated with either Alexa Fluor 488 or rhodamine. The corresponding images were merged (Overlay). Scale bar, 100 µm. (B and C) Anterior epithelial cells (B) and equatorial epithelial and nascent fiber cells (C). Scale bar, 20 µm. (D) Different depths of the lens regions from equator as illustrated in panels a (equator), b (cortical), and c (nucleus), Cx50 and integrin α6 were double immunostained (left), and the extent of colocalization was quantified and presented as colocalization coefficiency using ImageJ (right). Thick black outline indicates the lens capsule. Scale bar, 20 µm. Each data point in the graph represents an individual mouse in each group. The data are presented as mean ± SEM. ***, P < 0.001.

    Techniques Used:

    Hemichannel opening by FFSS is regulated by integrin α6β1. (A) Differentiated chick lens primary culture was subjected to FFSS or treated with β1 Ab or IgG control at 20 µg/ml for 1 h separately or in combination, followed by EtBr/FITC-dextran dye uptake assay (upper panel). Scale bar, 100 µm. FITC-dextran (M r 10 kD) was used as a control for nonspecific membrane permeability. The level of dye uptake was quantified by subtracting FITC-positive from EtBr-positive cells (lower panel). Each data point in the graph represents an individual lentoid in one of three independent experiments. (B) Differentiated chick lens primary culture was pretreated with integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h, and then subjected to FFSS, or was pretreated with β1 Ab at 20 µg/ml for 1 h, followed by EtBr/FITC-dextran dye uptake (upper panel). Scale bar, 100 µm. The level of EtBr dye uptake was quantified by subtracting FITC-dextran–positive from EtBr-positive cells (lower panel). Each data point in the graph represents an individual lentoid in one of three independent experiments. (C) CEF cells expressing exogenous Cx50 or Cx46 via recombinant RCAS(A) infection or RCAS(A) vehicle control (V) were mechanically stimulated by dropping the medium from a fixed distance or treated with β1 Ab at 20 µg/ml for 1 h. The percentage of LY dye uptake–positive cells per image was quantified by subtracting both LY- and RD-positive cells (M r 10 kD). Each data point in the graph represents an individual quantified image in one of three independent experiments. (D) CEF cells expressing exogenous Cx50 via recombinant RCAS(A) infection or RCAS(A) vehicle control (V) were mechanically stimulated by dropping the medium from a fixed distance or were pretreated with integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h. The percentage of LY dye uptake positive cells per image was quantified by subtracting both LY- and RD-positive cells. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. NS, not significant; **, P < 0.01; ***, P < 0.001.
    Figure Legend Snippet: Hemichannel opening by FFSS is regulated by integrin α6β1. (A) Differentiated chick lens primary culture was subjected to FFSS or treated with β1 Ab or IgG control at 20 µg/ml for 1 h separately or in combination, followed by EtBr/FITC-dextran dye uptake assay (upper panel). Scale bar, 100 µm. FITC-dextran (M r 10 kD) was used as a control for nonspecific membrane permeability. The level of dye uptake was quantified by subtracting FITC-positive from EtBr-positive cells (lower panel). Each data point in the graph represents an individual lentoid in one of three independent experiments. (B) Differentiated chick lens primary culture was pretreated with integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h, and then subjected to FFSS, or was pretreated with β1 Ab at 20 µg/ml for 1 h, followed by EtBr/FITC-dextran dye uptake (upper panel). Scale bar, 100 µm. The level of EtBr dye uptake was quantified by subtracting FITC-dextran–positive from EtBr-positive cells (lower panel). Each data point in the graph represents an individual lentoid in one of three independent experiments. (C) CEF cells expressing exogenous Cx50 or Cx46 via recombinant RCAS(A) infection or RCAS(A) vehicle control (V) were mechanically stimulated by dropping the medium from a fixed distance or treated with β1 Ab at 20 µg/ml for 1 h. The percentage of LY dye uptake–positive cells per image was quantified by subtracting both LY- and RD-positive cells (M r 10 kD). Each data point in the graph represents an individual quantified image in one of three independent experiments. (D) CEF cells expressing exogenous Cx50 via recombinant RCAS(A) infection or RCAS(A) vehicle control (V) were mechanically stimulated by dropping the medium from a fixed distance or were pretreated with integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h. The percentage of LY dye uptake positive cells per image was quantified by subtracting both LY- and RD-positive cells. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. NS, not significant; **, P < 0.01; ***, P < 0.001.

    Techniques Used: Control, Membrane, Permeability, Blocking Assay, Expressing, Recombinant, Infection

    Hemichannel opening by FFSS is regulated by integrin α6β1 in lens epithelial cells. Primary chick lens culture was pretreated with or without integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h, then subjected to FFSS at 1 dyn/cm 2 for 10 min or treated with β1 Ab at 20 µg/ml for 1 h separately or in combination, followed by EtBr/FITC dye uptake assay. FITC-dextran (M r 10 kD) was used as a control for nonspecific membrane permeability (dying cells). The monolayer epithelial cells were imaged, and the level of EtBr uptake was quantified with EtBr dye uptake (excluding cells that took up both EtBr and FITC-dextran) using ImageJ. Each data point in the graph represents an individual quantified epithelial cell in one of three independent experiments. The data are presented as mean ± SEM. ***, P < 0.001.
    Figure Legend Snippet: Hemichannel opening by FFSS is regulated by integrin α6β1 in lens epithelial cells. Primary chick lens culture was pretreated with or without integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h, then subjected to FFSS at 1 dyn/cm 2 for 10 min or treated with β1 Ab at 20 µg/ml for 1 h separately or in combination, followed by EtBr/FITC dye uptake assay. FITC-dextran (M r 10 kD) was used as a control for nonspecific membrane permeability (dying cells). The monolayer epithelial cells were imaged, and the level of EtBr uptake was quantified with EtBr dye uptake (excluding cells that took up both EtBr and FITC-dextran) using ImageJ. Each data point in the graph represents an individual quantified epithelial cell in one of three independent experiments. The data are presented as mean ± SEM. ***, P < 0.001.

    Techniques Used: Blocking Assay, Control, Membrane, Permeability

    Integrin α6 is required in mechanical loading–induced opening of heteromeric hemichannels formed by Cx50 and Cx46. (A) CEF cells were infected with recombinant RCAS(A) retroviruses containing Cx50 or Cx46 or coinfected with retroviruses containing Cx46 with Cx50 or Cx50 mutant P88S. Crude cell membrane extracts were prepared and immunoblotted with anti-FLAG-tag and β-actin antibodies. The relative ratio of band intensity of total connexins (Cx50 and/or Cx46) to housekeeping protein β-actin with the ratio of the control setting as 1 is shown underneath the immunoblot. (B) The coinfected CEF cells expressing both Cx50 and Cx46 were pretreated with α6 Ab, followed by fluid dropping mechanical stimulation. LY/RD dye uptake assay was conducted, and the percentage of cells with LY dye uptake (excluding cells that took up both LY and RD-dextran) was quantified. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. **, P < 0.01.
    Figure Legend Snippet: Integrin α6 is required in mechanical loading–induced opening of heteromeric hemichannels formed by Cx50 and Cx46. (A) CEF cells were infected with recombinant RCAS(A) retroviruses containing Cx50 or Cx46 or coinfected with retroviruses containing Cx46 with Cx50 or Cx50 mutant P88S. Crude cell membrane extracts were prepared and immunoblotted with anti-FLAG-tag and β-actin antibodies. The relative ratio of band intensity of total connexins (Cx50 and/or Cx46) to housekeeping protein β-actin with the ratio of the control setting as 1 is shown underneath the immunoblot. (B) The coinfected CEF cells expressing both Cx50 and Cx46 were pretreated with α6 Ab, followed by fluid dropping mechanical stimulation. LY/RD dye uptake assay was conducted, and the percentage of cells with LY dye uptake (excluding cells that took up both LY and RD-dextran) was quantified. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. **, P < 0.01.

    Techniques Used: Infection, Recombinant, Mutagenesis, Membrane, FLAG-tag, Control, Western Blot, Expressing

    Activation of integrin β1 opens heteromeric hemichannels formed by Cx50 and Cx46. (A) CEF cells expressing exogenous Cx46 and Cx50 or Cx50 mutant P88S separately or in combination were seeded at low density to minimize gap junction formation. LY/RD-dextran dye uptake was performed after incubation with control IgG (IgG) or β1 Ab at 20 µg/ml for 1 h. (B) The percentage of LY-positive cells per image was counted and quantified by subtracting RD-positive cells. Scale bar, 50 µm. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. ***, P < 0.001.
    Figure Legend Snippet: Activation of integrin β1 opens heteromeric hemichannels formed by Cx50 and Cx46. (A) CEF cells expressing exogenous Cx46 and Cx50 or Cx50 mutant P88S separately or in combination were seeded at low density to minimize gap junction formation. LY/RD-dextran dye uptake was performed after incubation with control IgG (IgG) or β1 Ab at 20 µg/ml for 1 h. (B) The percentage of LY-positive cells per image was counted and quantified by subtracting RD-positive cells. Scale bar, 50 µm. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. ***, P < 0.001.

    Techniques Used: Activation Assay, Expressing, Mutagenesis, Incubation, Control

    Integrin α6 associates with lens Cx50. (A) Crude membrane extracts of cortical fiber portions of E19 chick lens as illustrated in the diagram (left) were collected and immunoprecipitated with control IgG or anti-Cx50 CT antibody. Preloading lysate (Input) and immunoprecipitates (IP) were immunoblotted with anti-integrin α6 and anti-Cx50 loop domain antibody (right). (B) CEF cells infected with RCAS(A) Cx50 were subjected to FFSS at 1 dyn/cm 2 for 30 min (FFSS) or under static conditions (C) and immunoprecipitated with anti-FLAG antibody. The immunoprecipitates were immunoblotted with anti-integrin α6 and anti-Cx50 CT antibody (upper panel). Relative immunoprecipitated integrin α6 to immunoprecipitated Cx50 band intensity was quantified (lower panel). Each data point in the graph represents an individual experiment out of three repeats ( n = 3). (C) CEF cells infected with RCAS(A) Cx50 and Cx50 mutants 368T and 379T. Crude cell membrane extracts were immunoprecipitated with IgG control or anti-Cx50 intracellular loop domain antibody. The immunoprecipitates were immunoblotted with anti-integrin α6 or anti-Cx50 loop domain antibody (left). The truncation sites at C-terminus of Cx50 are indicated (right). Data are presented as mean ± SEM. *, P < 0.05.
    Figure Legend Snippet: Integrin α6 associates with lens Cx50. (A) Crude membrane extracts of cortical fiber portions of E19 chick lens as illustrated in the diagram (left) were collected and immunoprecipitated with control IgG or anti-Cx50 CT antibody. Preloading lysate (Input) and immunoprecipitates (IP) were immunoblotted with anti-integrin α6 and anti-Cx50 loop domain antibody (right). (B) CEF cells infected with RCAS(A) Cx50 were subjected to FFSS at 1 dyn/cm 2 for 30 min (FFSS) or under static conditions (C) and immunoprecipitated with anti-FLAG antibody. The immunoprecipitates were immunoblotted with anti-integrin α6 and anti-Cx50 CT antibody (upper panel). Relative immunoprecipitated integrin α6 to immunoprecipitated Cx50 band intensity was quantified (lower panel). Each data point in the graph represents an individual experiment out of three repeats ( n = 3). (C) CEF cells infected with RCAS(A) Cx50 and Cx50 mutants 368T and 379T. Crude cell membrane extracts were immunoprecipitated with IgG control or anti-Cx50 intracellular loop domain antibody. The immunoprecipitates were immunoblotted with anti-integrin α6 or anti-Cx50 loop domain antibody (left). The truncation sites at C-terminus of Cx50 are indicated (right). Data are presented as mean ± SEM. *, P < 0.05.

    Techniques Used: Membrane, Immunoprecipitation, Control, Infection

    Association of integrin α6 and Cx50 is essential for the permeability of glucose and GSH through hemichannels. (A) Crude membrane extracts from CEF cells infected with RCAS(A) vehicle (V), Cx50 WT, or Cx50 truncation mutants 368T or 379T were isolated and immunoblotted with anti-Cx50 loop antibody anti-β-actin antibody. The expression of retrovirus-induced exogenous connexins in the CEF cells was quantified. The relative ratio of band intensity of Cx50 or truncated mutants to β-actin with the ratio of the control setting as 1 is shown underneath the immunoblot. (B) CEF cells were infected with RCAS(A) containing Cx50 WT, Cx50(368T), Cx50(379T), or RCAS(A) vehicle control (V). Cx50 WT was pretreated with or without α6 blocking antibody at 10 µg/ml for 1 h, and then subjected to FFSS for 30 min or under static conditions. GSH uptake was conducted and quantified. Each data point in the graph represents an individual quantified image in one of three independent experiments. (C) CEF cells were infected with RCAS(A) containing Cx50 WT or mutants 368T and 379T, or RCAS(A) vehicle control (V). WT Cx50 was pretreated with or without anti-integrin α6 antibody at 10 µg/ml in glucose free medium for 1 h. Cells were then subjected to FFSS for 30 min or under static conditions. Glucose uptake was conducted and analyzed by flow cytometry (left). The mean fluorescent intensity of 2-NBDG uptake was quantified (right). Each data point in the graph represents an individual quantified image in one of three independent experiments. All data are presented as mean ± SEM. ***, P < 0.001.
    Figure Legend Snippet: Association of integrin α6 and Cx50 is essential for the permeability of glucose and GSH through hemichannels. (A) Crude membrane extracts from CEF cells infected with RCAS(A) vehicle (V), Cx50 WT, or Cx50 truncation mutants 368T or 379T were isolated and immunoblotted with anti-Cx50 loop antibody anti-β-actin antibody. The expression of retrovirus-induced exogenous connexins in the CEF cells was quantified. The relative ratio of band intensity of Cx50 or truncated mutants to β-actin with the ratio of the control setting as 1 is shown underneath the immunoblot. (B) CEF cells were infected with RCAS(A) containing Cx50 WT, Cx50(368T), Cx50(379T), or RCAS(A) vehicle control (V). Cx50 WT was pretreated with or without α6 blocking antibody at 10 µg/ml for 1 h, and then subjected to FFSS for 30 min or under static conditions. GSH uptake was conducted and quantified. Each data point in the graph represents an individual quantified image in one of three independent experiments. (C) CEF cells were infected with RCAS(A) containing Cx50 WT or mutants 368T and 379T, or RCAS(A) vehicle control (V). WT Cx50 was pretreated with or without anti-integrin α6 antibody at 10 µg/ml in glucose free medium for 1 h. Cells were then subjected to FFSS for 30 min or under static conditions. Glucose uptake was conducted and analyzed by flow cytometry (left). The mean fluorescent intensity of 2-NBDG uptake was quantified (right). Each data point in the graph represents an individual quantified image in one of three independent experiments. All data are presented as mean ± SEM. ***, P < 0.001.

    Techniques Used: Permeability, Membrane, Infection, Isolation, Expressing, Control, Western Blot, Blocking Assay, Flow Cytometry

    Integrin α6 is required for Cx50 hemichannel opening. (A) Different lengths of single lens fiber cells were isolated from WT mice and mechanically loaded by spinning force (SF; 1,000 rpm for 5 min). An EtBr dye uptake assay was performed. Scale bar, 50 µm (left). The morphology of unhealthy fiber cells resembles resealed globules or has a rounded appearance . Unhealthy or dead fibers are readily distinguished by their morphology without using FITC dextran. Isolated single fibers were grouped by their lengths; short fibers (S) were <200 µm long; medium fibers (M) were 200 to 400 µm; and long fibers (L) were >400 µm. EtBr dye uptake intensity was quantified by ImageJ (right). (B) Isolated single lens fibers were obtained from WT mouse lenses and immunostained with integrin α6 antibody. (C) Single lens fibers were isolated from WT, Cx46 KO, and Cx50 KO mouse lenses. The fibers were pretreated with or without anti-integrin α6 antibody (α6 Ab; 10 µg/ml for 20 min), and EtBr dye uptake assay was performed during mechanical stimulation with SF (1,000 rpm for 5 min). EtBr dye uptake intensity in short fibers was quantified. Each data point in the graph represents an individual single fiber in one of three independent experiments. All data are presented as mean ± SEM. NS, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
    Figure Legend Snippet: Integrin α6 is required for Cx50 hemichannel opening. (A) Different lengths of single lens fiber cells were isolated from WT mice and mechanically loaded by spinning force (SF; 1,000 rpm for 5 min). An EtBr dye uptake assay was performed. Scale bar, 50 µm (left). The morphology of unhealthy fiber cells resembles resealed globules or has a rounded appearance . Unhealthy or dead fibers are readily distinguished by their morphology without using FITC dextran. Isolated single fibers were grouped by their lengths; short fibers (S) were <200 µm long; medium fibers (M) were 200 to 400 µm; and long fibers (L) were >400 µm. EtBr dye uptake intensity was quantified by ImageJ (right). (B) Isolated single lens fibers were obtained from WT mouse lenses and immunostained with integrin α6 antibody. (C) Single lens fibers were isolated from WT, Cx46 KO, and Cx50 KO mouse lenses. The fibers were pretreated with or without anti-integrin α6 antibody (α6 Ab; 10 µg/ml for 20 min), and EtBr dye uptake assay was performed during mechanical stimulation with SF (1,000 rpm for 5 min). EtBr dye uptake intensity in short fibers was quantified. Each data point in the graph represents an individual single fiber in one of three independent experiments. All data are presented as mean ± SEM. NS, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

    Techniques Used: Isolation

    Diagram of the mechanism of connexin hemichannels activated by FFSS and the mechanism of glucose and GSH transport to achieve lens fiber homeostasis. (A) FFSS induced by flow of fluid in the microcirculation leads to conformational activation of integrin α6β1. Activated integrin α6β1 enhances the association with C-terminus of Cx50 (black tail) and facilitates connexin hemichannels opening and the transport of GLU and GSH into lens fiber cells. (B) Cx50 hemichannels act as a transport portal for the uptake of GLU and GSH into the outer cortical fibers when glucose and GSH concentrations in the extracellular space are higher than those of the intracellular space (1). Cx50 hemichannels could also release GSH by outer lens epithelium/immature fibers when GSH concentration is higher intracellularly than extracellularly (2). This process helps GSH diffuse to proximal lens fiber cells. Glucose might not be able to release from fiber cells because glucose turns to glucose-6-phosphate rapidly inside cells . Because of the physiological barrier , nutrients and antioxidants taking up from outer cortical fibers will be transported from outer lens to the nuclear fibers through gap junctions, likely by Cx46 . In the nuclear fibers, because of the absence of integrin α6β1 and truncation of Cx50 at its C-terminus, hemichannels formed by truncated Cx50 (green) are not responsive to mechanical stimulation and thus remain closed. In this diagram: HC, hemichannel; yellow, full-length HC; green, truncated HC; GJ, gap junction; GLU (red hexagon), glucose; red hexagon with green dots, glucose-6-phosphate.
    Figure Legend Snippet: Diagram of the mechanism of connexin hemichannels activated by FFSS and the mechanism of glucose and GSH transport to achieve lens fiber homeostasis. (A) FFSS induced by flow of fluid in the microcirculation leads to conformational activation of integrin α6β1. Activated integrin α6β1 enhances the association with C-terminus of Cx50 (black tail) and facilitates connexin hemichannels opening and the transport of GLU and GSH into lens fiber cells. (B) Cx50 hemichannels act as a transport portal for the uptake of GLU and GSH into the outer cortical fibers when glucose and GSH concentrations in the extracellular space are higher than those of the intracellular space (1). Cx50 hemichannels could also release GSH by outer lens epithelium/immature fibers when GSH concentration is higher intracellularly than extracellularly (2). This process helps GSH diffuse to proximal lens fiber cells. Glucose might not be able to release from fiber cells because glucose turns to glucose-6-phosphate rapidly inside cells . Because of the physiological barrier , nutrients and antioxidants taking up from outer cortical fibers will be transported from outer lens to the nuclear fibers through gap junctions, likely by Cx46 . In the nuclear fibers, because of the absence of integrin α6β1 and truncation of Cx50 at its C-terminus, hemichannels formed by truncated Cx50 (green) are not responsive to mechanical stimulation and thus remain closed. In this diagram: HC, hemichannel; yellow, full-length HC; green, truncated HC; GJ, gap junction; GLU (red hexagon), glucose; red hexagon with green dots, glucose-6-phosphate.

    Techniques Used: Activation Assay, Concentration Assay

    Related Articles

    Activation Assay:

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    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
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    Mutagenesis:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
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    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
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    Isolation:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
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    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Positive Control:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Knock-Out:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Labeling:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Confocal Microscopy:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Inhibition:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Expressing:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Activity Assay:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    TUNEL Assay:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Translocation Assay:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Immunoprecipitation:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Laser Capture Microdissection:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Control:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Knockdown:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Western Blot:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Immunostaining:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Staining:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Fluorescence:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Cell Differentiation:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Imaging:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.

    Blocking Assay:

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.. Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.Immunostaining Whole eyes isolated from α6 +/+ and α6 −/− mice were fixed in 3.7% formaldehyde solution for 18–24 h at 4 °C and then incubated in 30% sucrose solution prior to cryofreezing and cryosectioning.

    Article Title: α6 Integrin Transactivates Insulin-like Growth Factor Receptor-1 (IGF-1R) to Regulate Caspase-3-mediated Lens Epithelial Cell Differentiation Initiation
    Article Snippet: Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.Anti-Aquaporin-0 (AB3071) and 4G10 antibody (05-1050) were purchased from EMD Millipore.. P2C62C4 mouse monoclonal antibody to chicken α6 integrin was obtained from the Developmental Studies Hybridoma Bank.



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    integrin α 6  (Santa Cruz Biotechnology)
    88
    Santa Cruz Biotechnology integrin α 6
    <t>Integrin</t> <t>α6</t> is expressed in cortical lens, primary chick lens culture, and CEF cells. (A) Crude membrane extracts were prepared from whole chick lens, CEF cells, and differentiated primary chick lens culture containing lentoids and immunoblotted with <t>anti-integrin</t> <t>α6</t> antibody and β-actin antibody. (B) Crude membrane extracts were prepared from whole chick lens (W) or isolated E18 chick lens portions containing cortical fibers (CF) or nuclear fibers (NF). Integrin α6 was detected in the cortical lens fibers, but not in the central, nuclear lens fiber. As no actin was expressed in nuclear lens, Ponceau S staining was used to show that a comparable amount of protein was loaded on SDS-PAGE.
    Integrin α 6, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    chicken α6 integrin  (Developmental Studies Hybridoma Bank)
    95
    Developmental Studies Hybridoma Bank chicken α6 integrin
    <t>Integrin</t> <t>α6</t> is expressed in cortical lens, primary chick lens culture, and CEF cells. (A) Crude membrane extracts were prepared from whole chick lens, CEF cells, and differentiated primary chick lens culture containing lentoids and immunoblotted with <t>anti-integrin</t> <t>α6</t> antibody and β-actin antibody. (B) Crude membrane extracts were prepared from whole chick lens (W) or isolated E18 chick lens portions containing cortical fibers (CF) or nuclear fibers (NF). Integrin α6 was detected in the cortical lens fibers, but not in the central, nuclear lens fiber. As no actin was expressed in nuclear lens, Ponceau S staining was used to show that a comparable amount of protein was loaded on SDS-PAGE.
    Chicken α6 Integrin, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/chicken+%CE%B16+integrin/anti-Integrin/pmc03924254-161-5-12
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    chicken α6 integrin - by Bioz Stars, 2026-09
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    α6 integrin subunit  (Santa Cruz Biotechnology)
    88
    Santa Cruz Biotechnology α6 integrin subunit
    Figure 1.Netrin-4 binds to endothelial 61 <t>integrin.</t> A, Expression of different integrin subunits as well as laminin 1 was determined in human umbilical vein (HUVECs), umbilical artery (HUAECs), microvascular dermal blood (HMVEC-ds), and microvascular dermal lymphatic (HMVEC-dLys) endothelial cells by quantita- tive RT-PCR normalized to brain expression. B, (upper panel) Expression of <t>6</t> and 1 integrin subunits in HMVEC-dLys was con- firmed by Western blotting (WB). B, (lower panel) Determination of the 61 integrin heterodimer expression in HMVEC-dLys by sequential immunoprecipitation (IP) and West- ern blotting (IB) with antibodies anti-1 (P5D2, 9EG7, 6S6) and 6 integrin subunits, respec- tively. C, (left panel) Immunoprecipitation with anti 6 integrin subunit antibody or its isotype control, followed by Western blotting for netrin-4. Recombinant netrin-4 was used as a positive control (input) (MM molecular marker). (right panel) Pull-down between 6 or 1-GST recombinant fusion proteins in the absence or presence of netrin-4, followed by netrin-4 Western blotting. D, Binding of netrin-4 (0.001 to 0.5 g/mL) to microplate- coated proteins (10 g/mL each); laminin-111 (black), 6 (red), or v5 (blue) integrin was determined by ELISA. Three separate experi- ments were done in duplicate. E, Activation of 1 integrin subunit was determined by seed- ing HMVEC-dLys on control (buffer only), netrin-4-coated, or VEGF-C-coated (1 g/mL each) wells for 5 or 30 minutes, followed by Western blotting using the anti-1 integrin antibody (9EG7), which recognizes specifi- cally its active conformation. Total Src and coomassie blue staining of the membranes served as loading controls.
    α6 Integrin Subunit, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    α6 integrin subunit - by Bioz Stars, 2026-09
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    Image Search Results


    Integrin α6 is expressed in cortical lens, primary chick lens culture, and CEF cells. (A) Crude membrane extracts were prepared from whole chick lens, CEF cells, and differentiated primary chick lens culture containing lentoids and immunoblotted with anti-integrin α6 antibody and β-actin antibody. (B) Crude membrane extracts were prepared from whole chick lens (W) or isolated E18 chick lens portions containing cortical fibers (CF) or nuclear fibers (NF). Integrin α6 was detected in the cortical lens fibers, but not in the central, nuclear lens fiber. As no actin was expressed in nuclear lens, Ponceau S staining was used to show that a comparable amount of protein was loaded on SDS-PAGE.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Integrin α6 is expressed in cortical lens, primary chick lens culture, and CEF cells. (A) Crude membrane extracts were prepared from whole chick lens, CEF cells, and differentiated primary chick lens culture containing lentoids and immunoblotted with anti-integrin α6 antibody and β-actin antibody. (B) Crude membrane extracts were prepared from whole chick lens (W) or isolated E18 chick lens portions containing cortical fibers (CF) or nuclear fibers (NF). Integrin α6 was detected in the cortical lens fibers, but not in the central, nuclear lens fiber. As no actin was expressed in nuclear lens, Ponceau S staining was used to show that a comparable amount of protein was loaded on SDS-PAGE.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques: Membrane, Isolation, Staining, SDS Page

    Cx50 colocalized with integrin α6 in lens epithelium and outer cortical fiber cells. (A) Embryonic day 18 mouse lens sagittal cryosections were immunostained with anti-Cx46 (green), Cx50 (green), or integrin α6 (red) antibody and counterstained with DAPI (blue), followed by corresponding secondary antibodies conjugated with either Alexa Fluor 488 or rhodamine. The corresponding images were merged (Overlay). Scale bar, 100 µm. (B and C) Anterior epithelial cells (B) and equatorial epithelial and nascent fiber cells (C). Scale bar, 20 µm. (D) Different depths of the lens regions from equator as illustrated in panels a (equator), b (cortical), and c (nucleus), Cx50 and integrin α6 were double immunostained (left), and the extent of colocalization was quantified and presented as colocalization coefficiency using ImageJ (right). Thick black outline indicates the lens capsule. Scale bar, 20 µm. Each data point in the graph represents an individual mouse in each group. The data are presented as mean ± SEM. ***, P < 0.001.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Cx50 colocalized with integrin α6 in lens epithelium and outer cortical fiber cells. (A) Embryonic day 18 mouse lens sagittal cryosections were immunostained with anti-Cx46 (green), Cx50 (green), or integrin α6 (red) antibody and counterstained with DAPI (blue), followed by corresponding secondary antibodies conjugated with either Alexa Fluor 488 or rhodamine. The corresponding images were merged (Overlay). Scale bar, 100 µm. (B and C) Anterior epithelial cells (B) and equatorial epithelial and nascent fiber cells (C). Scale bar, 20 µm. (D) Different depths of the lens regions from equator as illustrated in panels a (equator), b (cortical), and c (nucleus), Cx50 and integrin α6 were double immunostained (left), and the extent of colocalization was quantified and presented as colocalization coefficiency using ImageJ (right). Thick black outline indicates the lens capsule. Scale bar, 20 µm. Each data point in the graph represents an individual mouse in each group. The data are presented as mean ± SEM. ***, P < 0.001.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques:

    Hemichannel opening by FFSS is regulated by integrin α6β1. (A) Differentiated chick lens primary culture was subjected to FFSS or treated with β1 Ab or IgG control at 20 µg/ml for 1 h separately or in combination, followed by EtBr/FITC-dextran dye uptake assay (upper panel). Scale bar, 100 µm. FITC-dextran (M r 10 kD) was used as a control for nonspecific membrane permeability. The level of dye uptake was quantified by subtracting FITC-positive from EtBr-positive cells (lower panel). Each data point in the graph represents an individual lentoid in one of three independent experiments. (B) Differentiated chick lens primary culture was pretreated with integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h, and then subjected to FFSS, or was pretreated with β1 Ab at 20 µg/ml for 1 h, followed by EtBr/FITC-dextran dye uptake (upper panel). Scale bar, 100 µm. The level of EtBr dye uptake was quantified by subtracting FITC-dextran–positive from EtBr-positive cells (lower panel). Each data point in the graph represents an individual lentoid in one of three independent experiments. (C) CEF cells expressing exogenous Cx50 or Cx46 via recombinant RCAS(A) infection or RCAS(A) vehicle control (V) were mechanically stimulated by dropping the medium from a fixed distance or treated with β1 Ab at 20 µg/ml for 1 h. The percentage of LY dye uptake–positive cells per image was quantified by subtracting both LY- and RD-positive cells (M r 10 kD). Each data point in the graph represents an individual quantified image in one of three independent experiments. (D) CEF cells expressing exogenous Cx50 via recombinant RCAS(A) infection or RCAS(A) vehicle control (V) were mechanically stimulated by dropping the medium from a fixed distance or were pretreated with integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h. The percentage of LY dye uptake positive cells per image was quantified by subtracting both LY- and RD-positive cells. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. NS, not significant; **, P < 0.01; ***, P < 0.001.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Hemichannel opening by FFSS is regulated by integrin α6β1. (A) Differentiated chick lens primary culture was subjected to FFSS or treated with β1 Ab or IgG control at 20 µg/ml for 1 h separately or in combination, followed by EtBr/FITC-dextran dye uptake assay (upper panel). Scale bar, 100 µm. FITC-dextran (M r 10 kD) was used as a control for nonspecific membrane permeability. The level of dye uptake was quantified by subtracting FITC-positive from EtBr-positive cells (lower panel). Each data point in the graph represents an individual lentoid in one of three independent experiments. (B) Differentiated chick lens primary culture was pretreated with integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h, and then subjected to FFSS, or was pretreated with β1 Ab at 20 µg/ml for 1 h, followed by EtBr/FITC-dextran dye uptake (upper panel). Scale bar, 100 µm. The level of EtBr dye uptake was quantified by subtracting FITC-dextran–positive from EtBr-positive cells (lower panel). Each data point in the graph represents an individual lentoid in one of three independent experiments. (C) CEF cells expressing exogenous Cx50 or Cx46 via recombinant RCAS(A) infection or RCAS(A) vehicle control (V) were mechanically stimulated by dropping the medium from a fixed distance or treated with β1 Ab at 20 µg/ml for 1 h. The percentage of LY dye uptake–positive cells per image was quantified by subtracting both LY- and RD-positive cells (M r 10 kD). Each data point in the graph represents an individual quantified image in one of three independent experiments. (D) CEF cells expressing exogenous Cx50 via recombinant RCAS(A) infection or RCAS(A) vehicle control (V) were mechanically stimulated by dropping the medium from a fixed distance or were pretreated with integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h. The percentage of LY dye uptake positive cells per image was quantified by subtracting both LY- and RD-positive cells. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. NS, not significant; **, P < 0.01; ***, P < 0.001.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques: Control, Membrane, Permeability, Blocking Assay, Expressing, Recombinant, Infection

    Hemichannel opening by FFSS is regulated by integrin α6β1 in lens epithelial cells. Primary chick lens culture was pretreated with or without integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h, then subjected to FFSS at 1 dyn/cm 2 for 10 min or treated with β1 Ab at 20 µg/ml for 1 h separately or in combination, followed by EtBr/FITC dye uptake assay. FITC-dextran (M r 10 kD) was used as a control for nonspecific membrane permeability (dying cells). The monolayer epithelial cells were imaged, and the level of EtBr uptake was quantified with EtBr dye uptake (excluding cells that took up both EtBr and FITC-dextran) using ImageJ. Each data point in the graph represents an individual quantified epithelial cell in one of three independent experiments. The data are presented as mean ± SEM. ***, P < 0.001.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Hemichannel opening by FFSS is regulated by integrin α6β1 in lens epithelial cells. Primary chick lens culture was pretreated with or without integrin α6 blocking antibody (α6 Ab) at 10 µg/ml for 1 h, then subjected to FFSS at 1 dyn/cm 2 for 10 min or treated with β1 Ab at 20 µg/ml for 1 h separately or in combination, followed by EtBr/FITC dye uptake assay. FITC-dextran (M r 10 kD) was used as a control for nonspecific membrane permeability (dying cells). The monolayer epithelial cells were imaged, and the level of EtBr uptake was quantified with EtBr dye uptake (excluding cells that took up both EtBr and FITC-dextran) using ImageJ. Each data point in the graph represents an individual quantified epithelial cell in one of three independent experiments. The data are presented as mean ± SEM. ***, P < 0.001.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques: Blocking Assay, Control, Membrane, Permeability

    Integrin α6 is required in mechanical loading–induced opening of heteromeric hemichannels formed by Cx50 and Cx46. (A) CEF cells were infected with recombinant RCAS(A) retroviruses containing Cx50 or Cx46 or coinfected with retroviruses containing Cx46 with Cx50 or Cx50 mutant P88S. Crude cell membrane extracts were prepared and immunoblotted with anti-FLAG-tag and β-actin antibodies. The relative ratio of band intensity of total connexins (Cx50 and/or Cx46) to housekeeping protein β-actin with the ratio of the control setting as 1 is shown underneath the immunoblot. (B) The coinfected CEF cells expressing both Cx50 and Cx46 were pretreated with α6 Ab, followed by fluid dropping mechanical stimulation. LY/RD dye uptake assay was conducted, and the percentage of cells with LY dye uptake (excluding cells that took up both LY and RD-dextran) was quantified. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. **, P < 0.01.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Integrin α6 is required in mechanical loading–induced opening of heteromeric hemichannels formed by Cx50 and Cx46. (A) CEF cells were infected with recombinant RCAS(A) retroviruses containing Cx50 or Cx46 or coinfected with retroviruses containing Cx46 with Cx50 or Cx50 mutant P88S. Crude cell membrane extracts were prepared and immunoblotted with anti-FLAG-tag and β-actin antibodies. The relative ratio of band intensity of total connexins (Cx50 and/or Cx46) to housekeeping protein β-actin with the ratio of the control setting as 1 is shown underneath the immunoblot. (B) The coinfected CEF cells expressing both Cx50 and Cx46 were pretreated with α6 Ab, followed by fluid dropping mechanical stimulation. LY/RD dye uptake assay was conducted, and the percentage of cells with LY dye uptake (excluding cells that took up both LY and RD-dextran) was quantified. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. **, P < 0.01.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques: Infection, Recombinant, Mutagenesis, Membrane, FLAG-tag, Control, Western Blot, Expressing

    Activation of integrin β1 opens heteromeric hemichannels formed by Cx50 and Cx46. (A) CEF cells expressing exogenous Cx46 and Cx50 or Cx50 mutant P88S separately or in combination were seeded at low density to minimize gap junction formation. LY/RD-dextran dye uptake was performed after incubation with control IgG (IgG) or β1 Ab at 20 µg/ml for 1 h. (B) The percentage of LY-positive cells per image was counted and quantified by subtracting RD-positive cells. Scale bar, 50 µm. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. ***, P < 0.001.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Activation of integrin β1 opens heteromeric hemichannels formed by Cx50 and Cx46. (A) CEF cells expressing exogenous Cx46 and Cx50 or Cx50 mutant P88S separately or in combination were seeded at low density to minimize gap junction formation. LY/RD-dextran dye uptake was performed after incubation with control IgG (IgG) or β1 Ab at 20 µg/ml for 1 h. (B) The percentage of LY-positive cells per image was counted and quantified by subtracting RD-positive cells. Scale bar, 50 µm. Each data point in the graph represents an individual quantified image in one of three independent experiments. The data are presented as mean ± SEM. ***, P < 0.001.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques: Activation Assay, Expressing, Mutagenesis, Incubation, Control

    Integrin α6 associates with lens Cx50. (A) Crude membrane extracts of cortical fiber portions of E19 chick lens as illustrated in the diagram (left) were collected and immunoprecipitated with control IgG or anti-Cx50 CT antibody. Preloading lysate (Input) and immunoprecipitates (IP) were immunoblotted with anti-integrin α6 and anti-Cx50 loop domain antibody (right). (B) CEF cells infected with RCAS(A) Cx50 were subjected to FFSS at 1 dyn/cm 2 for 30 min (FFSS) or under static conditions (C) and immunoprecipitated with anti-FLAG antibody. The immunoprecipitates were immunoblotted with anti-integrin α6 and anti-Cx50 CT antibody (upper panel). Relative immunoprecipitated integrin α6 to immunoprecipitated Cx50 band intensity was quantified (lower panel). Each data point in the graph represents an individual experiment out of three repeats ( n = 3). (C) CEF cells infected with RCAS(A) Cx50 and Cx50 mutants 368T and 379T. Crude cell membrane extracts were immunoprecipitated with IgG control or anti-Cx50 intracellular loop domain antibody. The immunoprecipitates were immunoblotted with anti-integrin α6 or anti-Cx50 loop domain antibody (left). The truncation sites at C-terminus of Cx50 are indicated (right). Data are presented as mean ± SEM. *, P < 0.05.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Integrin α6 associates with lens Cx50. (A) Crude membrane extracts of cortical fiber portions of E19 chick lens as illustrated in the diagram (left) were collected and immunoprecipitated with control IgG or anti-Cx50 CT antibody. Preloading lysate (Input) and immunoprecipitates (IP) were immunoblotted with anti-integrin α6 and anti-Cx50 loop domain antibody (right). (B) CEF cells infected with RCAS(A) Cx50 were subjected to FFSS at 1 dyn/cm 2 for 30 min (FFSS) or under static conditions (C) and immunoprecipitated with anti-FLAG antibody. The immunoprecipitates were immunoblotted with anti-integrin α6 and anti-Cx50 CT antibody (upper panel). Relative immunoprecipitated integrin α6 to immunoprecipitated Cx50 band intensity was quantified (lower panel). Each data point in the graph represents an individual experiment out of three repeats ( n = 3). (C) CEF cells infected with RCAS(A) Cx50 and Cx50 mutants 368T and 379T. Crude cell membrane extracts were immunoprecipitated with IgG control or anti-Cx50 intracellular loop domain antibody. The immunoprecipitates were immunoblotted with anti-integrin α6 or anti-Cx50 loop domain antibody (left). The truncation sites at C-terminus of Cx50 are indicated (right). Data are presented as mean ± SEM. *, P < 0.05.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques: Membrane, Immunoprecipitation, Control, Infection

    Association of integrin α6 and Cx50 is essential for the permeability of glucose and GSH through hemichannels. (A) Crude membrane extracts from CEF cells infected with RCAS(A) vehicle (V), Cx50 WT, or Cx50 truncation mutants 368T or 379T were isolated and immunoblotted with anti-Cx50 loop antibody anti-β-actin antibody. The expression of retrovirus-induced exogenous connexins in the CEF cells was quantified. The relative ratio of band intensity of Cx50 or truncated mutants to β-actin with the ratio of the control setting as 1 is shown underneath the immunoblot. (B) CEF cells were infected with RCAS(A) containing Cx50 WT, Cx50(368T), Cx50(379T), or RCAS(A) vehicle control (V). Cx50 WT was pretreated with or without α6 blocking antibody at 10 µg/ml for 1 h, and then subjected to FFSS for 30 min or under static conditions. GSH uptake was conducted and quantified. Each data point in the graph represents an individual quantified image in one of three independent experiments. (C) CEF cells were infected with RCAS(A) containing Cx50 WT or mutants 368T and 379T, or RCAS(A) vehicle control (V). WT Cx50 was pretreated with or without anti-integrin α6 antibody at 10 µg/ml in glucose free medium for 1 h. Cells were then subjected to FFSS for 30 min or under static conditions. Glucose uptake was conducted and analyzed by flow cytometry (left). The mean fluorescent intensity of 2-NBDG uptake was quantified (right). Each data point in the graph represents an individual quantified image in one of three independent experiments. All data are presented as mean ± SEM. ***, P < 0.001.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Association of integrin α6 and Cx50 is essential for the permeability of glucose and GSH through hemichannels. (A) Crude membrane extracts from CEF cells infected with RCAS(A) vehicle (V), Cx50 WT, or Cx50 truncation mutants 368T or 379T were isolated and immunoblotted with anti-Cx50 loop antibody anti-β-actin antibody. The expression of retrovirus-induced exogenous connexins in the CEF cells was quantified. The relative ratio of band intensity of Cx50 or truncated mutants to β-actin with the ratio of the control setting as 1 is shown underneath the immunoblot. (B) CEF cells were infected with RCAS(A) containing Cx50 WT, Cx50(368T), Cx50(379T), or RCAS(A) vehicle control (V). Cx50 WT was pretreated with or without α6 blocking antibody at 10 µg/ml for 1 h, and then subjected to FFSS for 30 min or under static conditions. GSH uptake was conducted and quantified. Each data point in the graph represents an individual quantified image in one of three independent experiments. (C) CEF cells were infected with RCAS(A) containing Cx50 WT or mutants 368T and 379T, or RCAS(A) vehicle control (V). WT Cx50 was pretreated with or without anti-integrin α6 antibody at 10 µg/ml in glucose free medium for 1 h. Cells were then subjected to FFSS for 30 min or under static conditions. Glucose uptake was conducted and analyzed by flow cytometry (left). The mean fluorescent intensity of 2-NBDG uptake was quantified (right). Each data point in the graph represents an individual quantified image in one of three independent experiments. All data are presented as mean ± SEM. ***, P < 0.001.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques: Permeability, Membrane, Infection, Isolation, Expressing, Control, Western Blot, Blocking Assay, Flow Cytometry

    Integrin α6 is required for Cx50 hemichannel opening. (A) Different lengths of single lens fiber cells were isolated from WT mice and mechanically loaded by spinning force (SF; 1,000 rpm for 5 min). An EtBr dye uptake assay was performed. Scale bar, 50 µm (left). The morphology of unhealthy fiber cells resembles resealed globules or has a rounded appearance . Unhealthy or dead fibers are readily distinguished by their morphology without using FITC dextran. Isolated single fibers were grouped by their lengths; short fibers (S) were <200 µm long; medium fibers (M) were 200 to 400 µm; and long fibers (L) were >400 µm. EtBr dye uptake intensity was quantified by ImageJ (right). (B) Isolated single lens fibers were obtained from WT mouse lenses and immunostained with integrin α6 antibody. (C) Single lens fibers were isolated from WT, Cx46 KO, and Cx50 KO mouse lenses. The fibers were pretreated with or without anti-integrin α6 antibody (α6 Ab; 10 µg/ml for 20 min), and EtBr dye uptake assay was performed during mechanical stimulation with SF (1,000 rpm for 5 min). EtBr dye uptake intensity in short fibers was quantified. Each data point in the graph represents an individual single fiber in one of three independent experiments. All data are presented as mean ± SEM. NS, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Integrin α6 is required for Cx50 hemichannel opening. (A) Different lengths of single lens fiber cells were isolated from WT mice and mechanically loaded by spinning force (SF; 1,000 rpm for 5 min). An EtBr dye uptake assay was performed. Scale bar, 50 µm (left). The morphology of unhealthy fiber cells resembles resealed globules or has a rounded appearance . Unhealthy or dead fibers are readily distinguished by their morphology without using FITC dextran. Isolated single fibers were grouped by their lengths; short fibers (S) were <200 µm long; medium fibers (M) were 200 to 400 µm; and long fibers (L) were >400 µm. EtBr dye uptake intensity was quantified by ImageJ (right). (B) Isolated single lens fibers were obtained from WT mouse lenses and immunostained with integrin α6 antibody. (C) Single lens fibers were isolated from WT, Cx46 KO, and Cx50 KO mouse lenses. The fibers were pretreated with or without anti-integrin α6 antibody (α6 Ab; 10 µg/ml for 20 min), and EtBr dye uptake assay was performed during mechanical stimulation with SF (1,000 rpm for 5 min). EtBr dye uptake intensity in short fibers was quantified. Each data point in the graph represents an individual single fiber in one of three independent experiments. All data are presented as mean ± SEM. NS, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques: Isolation

    Diagram of the mechanism of connexin hemichannels activated by FFSS and the mechanism of glucose and GSH transport to achieve lens fiber homeostasis. (A) FFSS induced by flow of fluid in the microcirculation leads to conformational activation of integrin α6β1. Activated integrin α6β1 enhances the association with C-terminus of Cx50 (black tail) and facilitates connexin hemichannels opening and the transport of GLU and GSH into lens fiber cells. (B) Cx50 hemichannels act as a transport portal for the uptake of GLU and GSH into the outer cortical fibers when glucose and GSH concentrations in the extracellular space are higher than those of the intracellular space (1). Cx50 hemichannels could also release GSH by outer lens epithelium/immature fibers when GSH concentration is higher intracellularly than extracellularly (2). This process helps GSH diffuse to proximal lens fiber cells. Glucose might not be able to release from fiber cells because glucose turns to glucose-6-phosphate rapidly inside cells . Because of the physiological barrier , nutrients and antioxidants taking up from outer cortical fibers will be transported from outer lens to the nuclear fibers through gap junctions, likely by Cx46 . In the nuclear fibers, because of the absence of integrin α6β1 and truncation of Cx50 at its C-terminus, hemichannels formed by truncated Cx50 (green) are not responsive to mechanical stimulation and thus remain closed. In this diagram: HC, hemichannel; yellow, full-length HC; green, truncated HC; GJ, gap junction; GLU (red hexagon), glucose; red hexagon with green dots, glucose-6-phosphate.

    Journal: The Journal of Cell Biology

    Article Title: Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens

    doi: 10.1083/jcb.202002154

    Figure Lengend Snippet: Diagram of the mechanism of connexin hemichannels activated by FFSS and the mechanism of glucose and GSH transport to achieve lens fiber homeostasis. (A) FFSS induced by flow of fluid in the microcirculation leads to conformational activation of integrin α6β1. Activated integrin α6β1 enhances the association with C-terminus of Cx50 (black tail) and facilitates connexin hemichannels opening and the transport of GLU and GSH into lens fiber cells. (B) Cx50 hemichannels act as a transport portal for the uptake of GLU and GSH into the outer cortical fibers when glucose and GSH concentrations in the extracellular space are higher than those of the intracellular space (1). Cx50 hemichannels could also release GSH by outer lens epithelium/immature fibers when GSH concentration is higher intracellularly than extracellularly (2). This process helps GSH diffuse to proximal lens fiber cells. Glucose might not be able to release from fiber cells because glucose turns to glucose-6-phosphate rapidly inside cells . Because of the physiological barrier , nutrients and antioxidants taking up from outer cortical fibers will be transported from outer lens to the nuclear fibers through gap junctions, likely by Cx46 . In the nuclear fibers, because of the absence of integrin α6β1 and truncation of Cx50 at its C-terminus, hemichannels formed by truncated Cx50 (green) are not responsive to mechanical stimulation and thus remain closed. In this diagram: HC, hemichannel; yellow, full-length HC; green, truncated HC; GJ, gap junction; GLU (red hexagon), glucose; red hexagon with green dots, glucose-6-phosphate.

    Article Snippet: Mouse anti-chicken integrin α6 for Western blot was obtained from the Developmental Studies Hybridoma Bank (P2C62C4; ).

    Techniques: Activation Assay, Concentration Assay

    Figure 1.Netrin-4 binds to endothelial 61 integrin. A, Expression of different integrin subunits as well as laminin 1 was determined in human umbilical vein (HUVECs), umbilical artery (HUAECs), microvascular dermal blood (HMVEC-ds), and microvascular dermal lymphatic (HMVEC-dLys) endothelial cells by quantita- tive RT-PCR normalized to brain expression. B, (upper panel) Expression of 6 and 1 integrin subunits in HMVEC-dLys was con- firmed by Western blotting (WB). B, (lower panel) Determination of the 61 integrin heterodimer expression in HMVEC-dLys by sequential immunoprecipitation (IP) and West- ern blotting (IB) with antibodies anti-1 (P5D2, 9EG7, 6S6) and 6 integrin subunits, respec- tively. C, (left panel) Immunoprecipitation with anti 6 integrin subunit antibody or its isotype control, followed by Western blotting for netrin-4. Recombinant netrin-4 was used as a positive control (input) (MM molecular marker). (right panel) Pull-down between 6 or 1-GST recombinant fusion proteins in the absence or presence of netrin-4, followed by netrin-4 Western blotting. D, Binding of netrin-4 (0.001 to 0.5 g/mL) to microplate- coated proteins (10 g/mL each); laminin-111 (black), 6 (red), or v5 (blue) integrin was determined by ELISA. Three separate experi- ments were done in duplicate. E, Activation of 1 integrin subunit was determined by seed- ing HMVEC-dLys on control (buffer only), netrin-4-coated, or VEGF-C-coated (1 g/mL each) wells for 5 or 30 minutes, followed by Western blotting using the anti-1 integrin antibody (9EG7), which recognizes specifi- cally its active conformation. Total Src and coomassie blue staining of the membranes served as loading controls.

    Journal: Circulation Research

    Article Title: Netrin-4 Activates Endothelial Integrin α6β1

    doi: 10.1161/circresaha.111.247239

    Figure Lengend Snippet: Figure 1.Netrin-4 binds to endothelial 61 integrin. A, Expression of different integrin subunits as well as laminin 1 was determined in human umbilical vein (HUVECs), umbilical artery (HUAECs), microvascular dermal blood (HMVEC-ds), and microvascular dermal lymphatic (HMVEC-dLys) endothelial cells by quantita- tive RT-PCR normalized to brain expression. B, (upper panel) Expression of 6 and 1 integrin subunits in HMVEC-dLys was con- firmed by Western blotting (WB). B, (lower panel) Determination of the 61 integrin heterodimer expression in HMVEC-dLys by sequential immunoprecipitation (IP) and West- ern blotting (IB) with antibodies anti-1 (P5D2, 9EG7, 6S6) and 6 integrin subunits, respec- tively. C, (left panel) Immunoprecipitation with anti 6 integrin subunit antibody or its isotype control, followed by Western blotting for netrin-4. Recombinant netrin-4 was used as a positive control (input) (MM molecular marker). (right panel) Pull-down between 6 or 1-GST recombinant fusion proteins in the absence or presence of netrin-4, followed by netrin-4 Western blotting. D, Binding of netrin-4 (0.001 to 0.5 g/mL) to microplate- coated proteins (10 g/mL each); laminin-111 (black), 6 (red), or v5 (blue) integrin was determined by ELISA. Three separate experi- ments were done in duplicate. E, Activation of 1 integrin subunit was determined by seed- ing HMVEC-dLys on control (buffer only), netrin-4-coated, or VEGF-C-coated (1 g/mL each) wells for 5 or 30 minutes, followed by Western blotting using the anti-1 integrin antibody (9EG7), which recognizes specifi- cally its active conformation. Total Src and coomassie blue staining of the membranes served as loading controls.

    Article Snippet: Equal amounts of proteins lysates were precleared with protein A/G Plus-agarose beads (Santa Cruz Biotechnology) and then combined with an anti α6 integrin subunit (N19, Santa Cruz Biotechnology), an anti β1 integrin subunit (P5B2, 9EG7 or 6S6, Millipore) or an isotype control (Santa Cruz Biotechnology), in presence or absence of 3 netrin-4 recombinant protein.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunoprecipitation, Control, Recombinant, Positive Control, Marker, Binding Assay, Enzyme-linked Immunosorbent Assay, Activation Assay, Staining

    Figure 2.Netrin-4-induced HMVEC-dLy migration and Src family kinase phosphorylation depends on 61 integrin. A, Che- motactic effects of netrin-4 (0.5 g/mL) on HMVEC-dLys in Boyden chamber assays in the absence () or presence of an isotype control (IgG), 6 (left panel, 10 to 40 g/mL) or 1 (right panel, 20 g/mL) function-blocking antibodies. B, HMVEC-dLys were transfected with anti-6 (left panel) or anti-1 integrin subunits (middle panel) siRNA. Silencing score was quantified by qRT- PCR and compared with control (Ctrl) siRNA transfected condition. Control or integrin subunits-transfected HMVEC-dLys under- went in vitro migration (right panel) stimulated by netrin-4 (0.5 g/mL). C, (upper panels) The 6 or 1 siRNA expression inhibits netrin-4-induced but not VEGF-C-induced (1 and 5 g/mL, respectively, for 15 minutes) SFK phosphorylation (Y416) as observed in control or 3 siRNA-transfected HMVEC-dLys. Inhibition of netrin-4-induced but not VEGF-C-induced (1 and 5 g/mL, for 15 minutes) SFK phosphorylation (Y416) is observed using 6 or 1, but not isotype control (IgG) or 3 function-blocking antibodies (40 g/mL) (lower panels). Experiments were at least performed in triplicate and change in phosphorylation (Y416) over total SFK determined using Image J and graphed as fold change over respective controls (right panels). *No growth factor control condi- tion. #Growth factor and antibody condition, P0.05.

    Journal: Circulation Research

    Article Title: Netrin-4 Activates Endothelial Integrin α6β1

    doi: 10.1161/circresaha.111.247239

    Figure Lengend Snippet: Figure 2.Netrin-4-induced HMVEC-dLy migration and Src family kinase phosphorylation depends on 61 integrin. A, Che- motactic effects of netrin-4 (0.5 g/mL) on HMVEC-dLys in Boyden chamber assays in the absence () or presence of an isotype control (IgG), 6 (left panel, 10 to 40 g/mL) or 1 (right panel, 20 g/mL) function-blocking antibodies. B, HMVEC-dLys were transfected with anti-6 (left panel) or anti-1 integrin subunits (middle panel) siRNA. Silencing score was quantified by qRT- PCR and compared with control (Ctrl) siRNA transfected condition. Control or integrin subunits-transfected HMVEC-dLys under- went in vitro migration (right panel) stimulated by netrin-4 (0.5 g/mL). C, (upper panels) The 6 or 1 siRNA expression inhibits netrin-4-induced but not VEGF-C-induced (1 and 5 g/mL, respectively, for 15 minutes) SFK phosphorylation (Y416) as observed in control or 3 siRNA-transfected HMVEC-dLys. Inhibition of netrin-4-induced but not VEGF-C-induced (1 and 5 g/mL, for 15 minutes) SFK phosphorylation (Y416) is observed using 6 or 1, but not isotype control (IgG) or 3 function-blocking antibodies (40 g/mL) (lower panels). Experiments were at least performed in triplicate and change in phosphorylation (Y416) over total SFK determined using Image J and graphed as fold change over respective controls (right panels). *No growth factor control condi- tion. #Growth factor and antibody condition, P0.05.

    Article Snippet: Equal amounts of proteins lysates were precleared with protein A/G Plus-agarose beads (Santa Cruz Biotechnology) and then combined with an anti α6 integrin subunit (N19, Santa Cruz Biotechnology), an anti β1 integrin subunit (P5B2, 9EG7 or 6S6, Millipore) or an isotype control (Santa Cruz Biotechnology), in presence or absence of 3 netrin-4 recombinant protein.

    Techniques: Migration, Phospho-proteomics, Control, Blocking Assay, Transfection, Quantitative RT-PCR, In Vitro, Expressing, Inhibition