mab3050 Search Results


human αvβ3 integrin  (R&D Systems)
92
R&D Systems human αvβ3 integrin
Immunostained slide with anti-avβ3 <t>integrin,</t> DAB chromogen, magnification x 200, score 0 in a case of unexplained infertility group
Human αvβ3 Integrin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human αvβ3 integrin/product/R&D Systems
Average 92 stars, based on 1 article reviews
human αvβ3 integrin - by Bioz Stars, 2026-03
92/100 stars
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lkb1  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc lkb1
<t>LKB1</t> controls apical junction formation in 16HBE cells. (A) At 2 days after infection with lentiviral vectors harboring control or 4 distinct shRNAs targeting LKB1 (shLKB1 to shLKB4), 16HBE cells were seeded on coverslips. At 2 days after plating, confluent cells were fixed and stained with anti-ZO-1 (top) and anti-E-cadherin (bottom). Bar, 20 μm. (B) (Top) Quantification of apical junction formation from three independent experiments. Error bar, SEMs. **, P < 0.01; ***, P < 0.001. (Bottom) At 5 days after infection, cells were lysed and analyzed by Western blotting with the indicated antibodies. Con, control. (C) (Top) Myc-tagged LKB1WT, LKB1KD (K78M), or LKB1SL26 was transiently transfected into 16HBE cells stably expressing HA-STRAD. One day later, cells were lysed and analyzed by Western blotting with the indicated antibodies. (Bottom) 16HBE cells stably expressing mLKB1WT, mLKB1KD (K78M), mLKB1SL26, or mLKB1C433A were infected with shRNA targeting LKB1 lentiviral vector, and 2 and 5 days later cells were lysed and analyzed by Western blotting with the indicated antibodies. (D) Cells described for panel C were replated, grown to confluence, and fixed and stained with anti-ZO-1 (bottom) and anti-myc (top). (E) Quantification of apical junction formation as described for panel D from three independent experiments. Error bars, SEMs. **, P < 0.01; ***, P < 0.001.
Lkb1, supplied by Cell Signaling Technology Inc, 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/result/lkb1/product/Cell Signaling Technology Inc
Average 95 stars, based on 1 article reviews
lkb1 - by Bioz Stars, 2026-03
95/100 stars
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itgavb3  (R&D Systems)
93
R&D Systems itgavb3
<t>LKB1</t> controls apical junction formation in 16HBE cells. (A) At 2 days after infection with lentiviral vectors harboring control or 4 distinct shRNAs targeting LKB1 (shLKB1 to shLKB4), 16HBE cells were seeded on coverslips. At 2 days after plating, confluent cells were fixed and stained with anti-ZO-1 (top) and anti-E-cadherin (bottom). Bar, 20 μm. (B) (Top) Quantification of apical junction formation from three independent experiments. Error bar, SEMs. **, P < 0.01; ***, P < 0.001. (Bottom) At 5 days after infection, cells were lysed and analyzed by Western blotting with the indicated antibodies. Con, control. (C) (Top) Myc-tagged LKB1WT, LKB1KD (K78M), or LKB1SL26 was transiently transfected into 16HBE cells stably expressing HA-STRAD. One day later, cells were lysed and analyzed by Western blotting with the indicated antibodies. (Bottom) 16HBE cells stably expressing mLKB1WT, mLKB1KD (K78M), mLKB1SL26, or mLKB1C433A were infected with shRNA targeting LKB1 lentiviral vector, and 2 and 5 days later cells were lysed and analyzed by Western blotting with the indicated antibodies. (D) Cells described for panel C were replated, grown to confluence, and fixed and stained with anti-ZO-1 (bottom) and anti-myc (top). (E) Quantification of apical junction formation as described for panel D from three independent experiments. Error bars, SEMs. **, P < 0.01; ***, P < 0.001.
Itgavb3, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/itgavb3/product/R&D Systems
Average 93 stars, based on 1 article reviews
itgavb3 - by Bioz Stars, 2026-03
93/100 stars
  Buy from Supplier

Image Search Results


Immunostained slide with anti-avβ3 integrin, DAB chromogen, magnification x 200, score 0 in a case of unexplained infertility group

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Immunostained slide with anti-avβ3 integrin, DAB chromogen, magnification x 200, score 0 in a case of unexplained infertility group

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Immunostained slide with anti-avβ3 integrin, DAB chromogen, magnification x 200, score 3 in a case of fertility group. Reactivity is mainly in the galndular epithelium)

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Immunostained slide with anti-avβ3 integrin, DAB chromogen, magnification x 200, score 3 in a case of fertility group. Reactivity is mainly in the galndular epithelium)

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Immunostained slide with anti- vβ3 integrin, DAB chromogen, magnification x 40 (Low Power), score 3 in A case of fertility group. Reactivity could be seen in both Luminal Epithelium (LE) the Glandular Epithelium (GE)

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Immunostained slide with anti- vβ3 integrin, DAB chromogen, magnification x 40 (Low Power), score 3 in A case of fertility group. Reactivity could be seen in both Luminal Epithelium (LE) the Glandular Epithelium (GE)

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Immunostained slide with anti- vβ3 integrin, DAB chromogen, magnification x 200 (High Power), score 3 in A case of fertility group. Reactivity could be seen Mainly in Luminal Epithelium (red arrow) rather than the Glandular Epithelium (red star)

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Immunostained slide with anti- vβ3 integrin, DAB chromogen, magnification x 200 (High Power), score 3 in A case of fertility group. Reactivity could be seen Mainly in Luminal Epithelium (red arrow) rather than the Glandular Epithelium (red star)

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Comparison of endometrial thickness, subendometrial Doppler resistance index, and ανβ3-integrin score in both study groups

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Comparison of endometrial thickness, subendometrial Doppler resistance index, and ανβ3-integrin score in both study groups

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques: Comparison, Control

Receiver-operating characteristic (ROC) curve for the discrimination between patients with unexplained infertility and normal controls using a : Endometrial thickness, b : Subendometril RI, c : avβ3 integrin

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Receiver-operating characteristic (ROC) curve for the discrimination between patients with unexplained infertility and normal controls using a : Endometrial thickness, b : Subendometril RI, c : avβ3 integrin

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Receiver-operating characteristic (ROC) curve analysis for the discrimination between patients with unexplained infertility and control group

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Receiver-operating characteristic (ROC) curve analysis for the discrimination between patients with unexplained infertility and control group

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques: Control

Summary of studies looked at endometrial αVβ3 integrin in luteal phase of infertile women

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Summary of studies looked at endometrial αVβ3 integrin in luteal phase of infertile women

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques: Control, Expressing

LKB1 controls apical junction formation in 16HBE cells. (A) At 2 days after infection with lentiviral vectors harboring control or 4 distinct shRNAs targeting LKB1 (shLKB1 to shLKB4), 16HBE cells were seeded on coverslips. At 2 days after plating, confluent cells were fixed and stained with anti-ZO-1 (top) and anti-E-cadherin (bottom). Bar, 20 μm. (B) (Top) Quantification of apical junction formation from three independent experiments. Error bar, SEMs. **, P < 0.01; ***, P < 0.001. (Bottom) At 5 days after infection, cells were lysed and analyzed by Western blotting with the indicated antibodies. Con, control. (C) (Top) Myc-tagged LKB1WT, LKB1KD (K78M), or LKB1SL26 was transiently transfected into 16HBE cells stably expressing HA-STRAD. One day later, cells were lysed and analyzed by Western blotting with the indicated antibodies. (Bottom) 16HBE cells stably expressing mLKB1WT, mLKB1KD (K78M), mLKB1SL26, or mLKB1C433A were infected with shRNA targeting LKB1 lentiviral vector, and 2 and 5 days later cells were lysed and analyzed by Western blotting with the indicated antibodies. (D) Cells described for panel C were replated, grown to confluence, and fixed and stained with anti-ZO-1 (bottom) and anti-myc (top). (E) Quantification of apical junction formation as described for panel D from three independent experiments. Error bars, SEMs. **, P < 0.01; ***, P < 0.001.

Journal: Molecular and Cellular Biology

Article Title: LKB1 Controls Human Bronchial Epithelial Morphogenesis through p114RhoGEF-Dependent RhoA Activation

doi: 10.1128/MCB.00154-13

Figure Lengend Snippet: LKB1 controls apical junction formation in 16HBE cells. (A) At 2 days after infection with lentiviral vectors harboring control or 4 distinct shRNAs targeting LKB1 (shLKB1 to shLKB4), 16HBE cells were seeded on coverslips. At 2 days after plating, confluent cells were fixed and stained with anti-ZO-1 (top) and anti-E-cadherin (bottom). Bar, 20 μm. (B) (Top) Quantification of apical junction formation from three independent experiments. Error bar, SEMs. **, P < 0.01; ***, P < 0.001. (Bottom) At 5 days after infection, cells were lysed and analyzed by Western blotting with the indicated antibodies. Con, control. (C) (Top) Myc-tagged LKB1WT, LKB1KD (K78M), or LKB1SL26 was transiently transfected into 16HBE cells stably expressing HA-STRAD. One day later, cells were lysed and analyzed by Western blotting with the indicated antibodies. (Bottom) 16HBE cells stably expressing mLKB1WT, mLKB1KD (K78M), mLKB1SL26, or mLKB1C433A were infected with shRNA targeting LKB1 lentiviral vector, and 2 and 5 days later cells were lysed and analyzed by Western blotting with the indicated antibodies. (D) Cells described for panel C were replated, grown to confluence, and fixed and stained with anti-ZO-1 (bottom) and anti-myc (top). (E) Quantification of apical junction formation as described for panel D from three independent experiments. Error bars, SEMs. **, P < 0.01; ***, P < 0.001.

Article Snippet: Primary antibodies used were ZO-1 (clone 1A12; 339100; Invitrogen, Carlsbad, CA), ZO-1 (617300; Invitrogen, Carlsbad, CA) E-cadherin (clone ECCD-2; 131900; Invitrogen, Carlsbad, CA), E-cadherin (clone 34; 610404; BD Transduction, Lexington, KY), α-tubulin (clone YL1/2; AbD Serotec, Raleigh, NC), hemagglutinin (HA; clone 3F10; Roche, Indianapolis, IN), green fluorescent protein (GFP; rabbit polyclonal; Invitrogen, Carlsbad, CA), myc (clone 9E10; Cancer Research UK, London, United Kingdom), phospho-PRK (p-PRK; 2611; Cell Signaling, Beverly, MA), PAK4 (3242; Cell Signaling, Beverly, MA), p114RhoGEF (EB06163; Everest Biotech Ramona, CA), LKB1 (27D10; Cell Signaling, Beverly, MA), and RhoA (sc-418; Santa Cruz, Santa Cruz, CA).

Techniques: Infection, Control, Staining, Western Blot, Transfection, Stable Transfection, Expressing, shRNA, Plasmid Preparation

LKB1 and STRAD activate RhoA and enhance phospho-PRK recruitment to apical junctions. (A) 16HBE cells stably expressing HA-tagged STRAD were infected with pLL3.7 lentiviral vectors encoding GFP-tagged LKB1 constructs. Twenty-four hours later, cells were harvested and Rho activity (Rho.GTP) was determined using a standard pulldown assay. LKB1WT, LKB1KD (K78M), and LKB1P38A induced 6- to 8-fold increases in the levels of active RhoA.GTP relative to the level for the control, LKB1SL26, or LKB1SL26/P38A. Total phospho-PRK was determined in the input cell lysates. Error bars, SEMs. **, P < 0.01; *, P < 0.05. WB, Western blotting. (B) 16HBE cells expressing GFP-tagged hLKB1P38A and hLKB1SL26/P38A were fixed and stained with DAPI (4′,6-diamidino-2-phenylindole). GFP signals were visualized directly. Bar, 20 μm. (C) At 1 day after infection, cells were stained for anti-phospho-PRK. GFP signals were visualized directly. GFP indicated infected cells. Bar, 20 μm.

Journal: Molecular and Cellular Biology

Article Title: LKB1 Controls Human Bronchial Epithelial Morphogenesis through p114RhoGEF-Dependent RhoA Activation

doi: 10.1128/MCB.00154-13

Figure Lengend Snippet: LKB1 and STRAD activate RhoA and enhance phospho-PRK recruitment to apical junctions. (A) 16HBE cells stably expressing HA-tagged STRAD were infected with pLL3.7 lentiviral vectors encoding GFP-tagged LKB1 constructs. Twenty-four hours later, cells were harvested and Rho activity (Rho.GTP) was determined using a standard pulldown assay. LKB1WT, LKB1KD (K78M), and LKB1P38A induced 6- to 8-fold increases in the levels of active RhoA.GTP relative to the level for the control, LKB1SL26, or LKB1SL26/P38A. Total phospho-PRK was determined in the input cell lysates. Error bars, SEMs. **, P < 0.01; *, P < 0.05. WB, Western blotting. (B) 16HBE cells expressing GFP-tagged hLKB1P38A and hLKB1SL26/P38A were fixed and stained with DAPI (4′,6-diamidino-2-phenylindole). GFP signals were visualized directly. Bar, 20 μm. (C) At 1 day after infection, cells were stained for anti-phospho-PRK. GFP signals were visualized directly. GFP indicated infected cells. Bar, 20 μm.

Article Snippet: Primary antibodies used were ZO-1 (clone 1A12; 339100; Invitrogen, Carlsbad, CA), ZO-1 (617300; Invitrogen, Carlsbad, CA) E-cadherin (clone ECCD-2; 131900; Invitrogen, Carlsbad, CA), E-cadherin (clone 34; 610404; BD Transduction, Lexington, KY), α-tubulin (clone YL1/2; AbD Serotec, Raleigh, NC), hemagglutinin (HA; clone 3F10; Roche, Indianapolis, IN), green fluorescent protein (GFP; rabbit polyclonal; Invitrogen, Carlsbad, CA), myc (clone 9E10; Cancer Research UK, London, United Kingdom), phospho-PRK (p-PRK; 2611; Cell Signaling, Beverly, MA), PAK4 (3242; Cell Signaling, Beverly, MA), p114RhoGEF (EB06163; Everest Biotech Ramona, CA), LKB1 (27D10; Cell Signaling, Beverly, MA), and RhoA (sc-418; Santa Cruz, Santa Cruz, CA).

Techniques: Stable Transfection, Expressing, Infection, Construct, Activity Assay, Control, Western Blot, Staining

p114RhoGEF controls apical junction formation in 16HBE cells and interacts with LKB1. (A) At 2 days after infection with lentiviral vectors harboring shRNAs targeting p114RhoGEF, 16HBE cells were seeded on coverslips. Two days later, confluent cells were fixed and stained with anti-ZO-1 (top) and anti-E-cadherin (bottom). Bar, 20 μm. (B) (Top) Quantification of apical junction formation from three independent experiments. Error bars, SEMs. *, P < 0.05; **, P < 0.01. (Bottom) At 5 days after infection, cells were lysed and analyzed by Western blotting with the indicated antibodies. (C) Schematic organization of hLKB1 and mouse p114RhoGEF (numbers represent amino acids). hLKB1 possesses a farnesylation site at aa 430. SL26 represents a 9-bp, in-frame deletion in the kinase domain. p114RhoGEF has a potential PBM at its C terminus. NRD, N-terminal regulatory domain; DH, Dbl homology domain. (D to F) HEK293T cells cotransfected with the indicated combinations of constructs. Proteins were immunoprecipitated (IP) from cell lysates using anti-mouse IgG or anti-Flag antibody. Input and immunoprecipitated lysates were analyzed by Western blotting.

Journal: Molecular and Cellular Biology

Article Title: LKB1 Controls Human Bronchial Epithelial Morphogenesis through p114RhoGEF-Dependent RhoA Activation

doi: 10.1128/MCB.00154-13

Figure Lengend Snippet: p114RhoGEF controls apical junction formation in 16HBE cells and interacts with LKB1. (A) At 2 days after infection with lentiviral vectors harboring shRNAs targeting p114RhoGEF, 16HBE cells were seeded on coverslips. Two days later, confluent cells were fixed and stained with anti-ZO-1 (top) and anti-E-cadherin (bottom). Bar, 20 μm. (B) (Top) Quantification of apical junction formation from three independent experiments. Error bars, SEMs. *, P < 0.05; **, P < 0.01. (Bottom) At 5 days after infection, cells were lysed and analyzed by Western blotting with the indicated antibodies. (C) Schematic organization of hLKB1 and mouse p114RhoGEF (numbers represent amino acids). hLKB1 possesses a farnesylation site at aa 430. SL26 represents a 9-bp, in-frame deletion in the kinase domain. p114RhoGEF has a potential PBM at its C terminus. NRD, N-terminal regulatory domain; DH, Dbl homology domain. (D to F) HEK293T cells cotransfected with the indicated combinations of constructs. Proteins were immunoprecipitated (IP) from cell lysates using anti-mouse IgG or anti-Flag antibody. Input and immunoprecipitated lysates were analyzed by Western blotting.

Article Snippet: Primary antibodies used were ZO-1 (clone 1A12; 339100; Invitrogen, Carlsbad, CA), ZO-1 (617300; Invitrogen, Carlsbad, CA) E-cadherin (clone ECCD-2; 131900; Invitrogen, Carlsbad, CA), E-cadherin (clone 34; 610404; BD Transduction, Lexington, KY), α-tubulin (clone YL1/2; AbD Serotec, Raleigh, NC), hemagglutinin (HA; clone 3F10; Roche, Indianapolis, IN), green fluorescent protein (GFP; rabbit polyclonal; Invitrogen, Carlsbad, CA), myc (clone 9E10; Cancer Research UK, London, United Kingdom), phospho-PRK (p-PRK; 2611; Cell Signaling, Beverly, MA), PAK4 (3242; Cell Signaling, Beverly, MA), p114RhoGEF (EB06163; Everest Biotech Ramona, CA), LKB1 (27D10; Cell Signaling, Beverly, MA), and RhoA (sc-418; Santa Cruz, Santa Cruz, CA).

Techniques: Infection, Staining, Western Blot, Construct, Immunoprecipitation

LKB1 and p114RhoGEF C-terminal domains have a dominant negative effect on apical junction formation in 16HBE cells. (A) 16HBE cells stably expressing GFP-tagged LKB1WT, LKB1KD, LKB1SL26, LKB1CRD (aa 309 to 433), or LKB1CRDΔCAAX (aa 309 to 429) were stained with anti-ZO-1 (bottom) and visualized directly with GFP signaling (top). (B) 16HBE cells stably expressing HA-tagged p114RhoGEF-FL (p114-FL), p114RhoGEF-N (p114-N), p114RhoGEF-M (p114-M), p114RhoGEF-C (p114-C), or p114RhoGEF-CΔPBM (p114-CΔPBM) were stained with anti-ZO-1 and anti-HA antibodies. Bars, 20 μm.

Journal: Molecular and Cellular Biology

Article Title: LKB1 Controls Human Bronchial Epithelial Morphogenesis through p114RhoGEF-Dependent RhoA Activation

doi: 10.1128/MCB.00154-13

Figure Lengend Snippet: LKB1 and p114RhoGEF C-terminal domains have a dominant negative effect on apical junction formation in 16HBE cells. (A) 16HBE cells stably expressing GFP-tagged LKB1WT, LKB1KD, LKB1SL26, LKB1CRD (aa 309 to 433), or LKB1CRDΔCAAX (aa 309 to 429) were stained with anti-ZO-1 (bottom) and visualized directly with GFP signaling (top). (B) 16HBE cells stably expressing HA-tagged p114RhoGEF-FL (p114-FL), p114RhoGEF-N (p114-N), p114RhoGEF-M (p114-M), p114RhoGEF-C (p114-C), or p114RhoGEF-CΔPBM (p114-CΔPBM) were stained with anti-ZO-1 and anti-HA antibodies. Bars, 20 μm.

Article Snippet: Primary antibodies used were ZO-1 (clone 1A12; 339100; Invitrogen, Carlsbad, CA), ZO-1 (617300; Invitrogen, Carlsbad, CA) E-cadherin (clone ECCD-2; 131900; Invitrogen, Carlsbad, CA), E-cadherin (clone 34; 610404; BD Transduction, Lexington, KY), α-tubulin (clone YL1/2; AbD Serotec, Raleigh, NC), hemagglutinin (HA; clone 3F10; Roche, Indianapolis, IN), green fluorescent protein (GFP; rabbit polyclonal; Invitrogen, Carlsbad, CA), myc (clone 9E10; Cancer Research UK, London, United Kingdom), phospho-PRK (p-PRK; 2611; Cell Signaling, Beverly, MA), PAK4 (3242; Cell Signaling, Beverly, MA), p114RhoGEF (EB06163; Everest Biotech Ramona, CA), LKB1 (27D10; Cell Signaling, Beverly, MA), and RhoA (sc-418; Santa Cruz, Santa Cruz, CA).

Techniques: Dominant Negative Mutation, Stable Transfection, Expressing, Staining

PLA to visualize the interaction between LKB1 with p114RhoGEF in 16HBE cells. (A) 16HBE cells stably expressing HA-tagged p114RhoGEF were fixed and stained with a mixture of mouse (Ms) anti-HA and rabbit (Rb) anti-LKB1 (A1), with anti-HA alone (A2), or with anti-LKB1 alone (A3). PLA signals were visualized by addition of a mixture of anti-mouse and anti-rabbit antibody reagents provided by the company (see Materials and Methods). Bar, 20 μm. (B) 16HBE cells stably expressing HA-tagged p114RhoGEF and either myc-LKB1WT (B1) or myc-LKB1SL26 (B2) were fixed and stained with a mixture of mouse anti-HA and rabbit anti-myc antibodies (left). PLA signals were visualized by addition of a mixture of anti-mouse and anti-rabbit antibody reagents provided by the company (see Materials and Methods). (C) Quantification of PLA signals. Data represent the number of positive dots per cell. Six random nonoverlapping images were taken, and signals were quantified using Volocity image analysis software. Error bars, SEMs. **, P < 0.01. (D) Fluorescent images of stable cells expressing HA-p114RhoGEF and myc-mLKB1WT stained with anti-HA antibody (left) and anti-myc antibody (right). Bar, 20 μm. (E) Fluorescent images of stable cells expressing HA-p114RhoGEF and myc-mLKB1SL26 stained with anti-HA antibody (left) and anti-myc antibody (right). (F) Lysates from 16HBE cells stably expressing HA-tagged p114RhoGEF and control cells were examined on Western blots with anti-p114RhoGEF or anti-LKB1 antibodies.

Journal: Molecular and Cellular Biology

Article Title: LKB1 Controls Human Bronchial Epithelial Morphogenesis through p114RhoGEF-Dependent RhoA Activation

doi: 10.1128/MCB.00154-13

Figure Lengend Snippet: PLA to visualize the interaction between LKB1 with p114RhoGEF in 16HBE cells. (A) 16HBE cells stably expressing HA-tagged p114RhoGEF were fixed and stained with a mixture of mouse (Ms) anti-HA and rabbit (Rb) anti-LKB1 (A1), with anti-HA alone (A2), or with anti-LKB1 alone (A3). PLA signals were visualized by addition of a mixture of anti-mouse and anti-rabbit antibody reagents provided by the company (see Materials and Methods). Bar, 20 μm. (B) 16HBE cells stably expressing HA-tagged p114RhoGEF and either myc-LKB1WT (B1) or myc-LKB1SL26 (B2) were fixed and stained with a mixture of mouse anti-HA and rabbit anti-myc antibodies (left). PLA signals were visualized by addition of a mixture of anti-mouse and anti-rabbit antibody reagents provided by the company (see Materials and Methods). (C) Quantification of PLA signals. Data represent the number of positive dots per cell. Six random nonoverlapping images were taken, and signals were quantified using Volocity image analysis software. Error bars, SEMs. **, P < 0.01. (D) Fluorescent images of stable cells expressing HA-p114RhoGEF and myc-mLKB1WT stained with anti-HA antibody (left) and anti-myc antibody (right). Bar, 20 μm. (E) Fluorescent images of stable cells expressing HA-p114RhoGEF and myc-mLKB1SL26 stained with anti-HA antibody (left) and anti-myc antibody (right). (F) Lysates from 16HBE cells stably expressing HA-tagged p114RhoGEF and control cells were examined on Western blots with anti-p114RhoGEF or anti-LKB1 antibodies.

Article Snippet: Primary antibodies used were ZO-1 (clone 1A12; 339100; Invitrogen, Carlsbad, CA), ZO-1 (617300; Invitrogen, Carlsbad, CA) E-cadherin (clone ECCD-2; 131900; Invitrogen, Carlsbad, CA), E-cadherin (clone 34; 610404; BD Transduction, Lexington, KY), α-tubulin (clone YL1/2; AbD Serotec, Raleigh, NC), hemagglutinin (HA; clone 3F10; Roche, Indianapolis, IN), green fluorescent protein (GFP; rabbit polyclonal; Invitrogen, Carlsbad, CA), myc (clone 9E10; Cancer Research UK, London, United Kingdom), phospho-PRK (p-PRK; 2611; Cell Signaling, Beverly, MA), PAK4 (3242; Cell Signaling, Beverly, MA), p114RhoGEF (EB06163; Everest Biotech Ramona, CA), LKB1 (27D10; Cell Signaling, Beverly, MA), and RhoA (sc-418; Santa Cruz, Santa Cruz, CA).

Techniques: Stable Transfection, Expressing, Staining, Software, Control, Western Blot

p114RhoGEF is required for LKB1-dependent RhoA activation. (Left) 16HBE cells stably expressing HA-tagged STRAD cells were transfected with siRNA control (siCon) or siRNA oligonucleotide targeting p114RhoGEF. Three days later, cells were infected with lentiviral vectors encoding LKB1WT, LKB1KD, and LKB1SL26. Twenty-four hours later, cells were harvested and the levels of activated Rho (Rho.GTP) were determined using a pulldown assay, followed by Western blotting. (Right) Quantification of the data from three experiments is shown. Error bars, SEMs. **, P < 0.01.

Journal: Molecular and Cellular Biology

Article Title: LKB1 Controls Human Bronchial Epithelial Morphogenesis through p114RhoGEF-Dependent RhoA Activation

doi: 10.1128/MCB.00154-13

Figure Lengend Snippet: p114RhoGEF is required for LKB1-dependent RhoA activation. (Left) 16HBE cells stably expressing HA-tagged STRAD cells were transfected with siRNA control (siCon) or siRNA oligonucleotide targeting p114RhoGEF. Three days later, cells were infected with lentiviral vectors encoding LKB1WT, LKB1KD, and LKB1SL26. Twenty-four hours later, cells were harvested and the levels of activated Rho (Rho.GTP) were determined using a pulldown assay, followed by Western blotting. (Right) Quantification of the data from three experiments is shown. Error bars, SEMs. **, P < 0.01.

Article Snippet: Primary antibodies used were ZO-1 (clone 1A12; 339100; Invitrogen, Carlsbad, CA), ZO-1 (617300; Invitrogen, Carlsbad, CA) E-cadherin (clone ECCD-2; 131900; Invitrogen, Carlsbad, CA), E-cadherin (clone 34; 610404; BD Transduction, Lexington, KY), α-tubulin (clone YL1/2; AbD Serotec, Raleigh, NC), hemagglutinin (HA; clone 3F10; Roche, Indianapolis, IN), green fluorescent protein (GFP; rabbit polyclonal; Invitrogen, Carlsbad, CA), myc (clone 9E10; Cancer Research UK, London, United Kingdom), phospho-PRK (p-PRK; 2611; Cell Signaling, Beverly, MA), PAK4 (3242; Cell Signaling, Beverly, MA), p114RhoGEF (EB06163; Everest Biotech Ramona, CA), LKB1 (27D10; Cell Signaling, Beverly, MA), and RhoA (sc-418; Santa Cruz, Santa Cruz, CA).

Techniques: Activation Assay, Stable Transfection, Expressing, Transfection, Control, Infection, Western Blot

LKB1 and p114RhoGEF regulate the maturation of primordial junctions to apical junctions. 16HBE cells infected with lentiviral vectors harboring control shRNA (A), shRNA2 targeting LKB1 (B), and shRNA3 targeting p114RhoGEF (C) were tested. At 5 days after infection, confluent monolayers were subjected to a calcium-switch assay. After calcium readdition, cells were fixed at 1 h and 6 h and stained with anti-ZO-1 (top) and anti-E-cadherin (bottom). Bar, 20 μm.

Journal: Molecular and Cellular Biology

Article Title: LKB1 Controls Human Bronchial Epithelial Morphogenesis through p114RhoGEF-Dependent RhoA Activation

doi: 10.1128/MCB.00154-13

Figure Lengend Snippet: LKB1 and p114RhoGEF regulate the maturation of primordial junctions to apical junctions. 16HBE cells infected with lentiviral vectors harboring control shRNA (A), shRNA2 targeting LKB1 (B), and shRNA3 targeting p114RhoGEF (C) were tested. At 5 days after infection, confluent monolayers were subjected to a calcium-switch assay. After calcium readdition, cells were fixed at 1 h and 6 h and stained with anti-ZO-1 (top) and anti-E-cadherin (bottom). Bar, 20 μm.

Article Snippet: Primary antibodies used were ZO-1 (clone 1A12; 339100; Invitrogen, Carlsbad, CA), ZO-1 (617300; Invitrogen, Carlsbad, CA) E-cadherin (clone ECCD-2; 131900; Invitrogen, Carlsbad, CA), E-cadherin (clone 34; 610404; BD Transduction, Lexington, KY), α-tubulin (clone YL1/2; AbD Serotec, Raleigh, NC), hemagglutinin (HA; clone 3F10; Roche, Indianapolis, IN), green fluorescent protein (GFP; rabbit polyclonal; Invitrogen, Carlsbad, CA), myc (clone 9E10; Cancer Research UK, London, United Kingdom), phospho-PRK (p-PRK; 2611; Cell Signaling, Beverly, MA), PAK4 (3242; Cell Signaling, Beverly, MA), p114RhoGEF (EB06163; Everest Biotech Ramona, CA), LKB1 (27D10; Cell Signaling, Beverly, MA), and RhoA (sc-418; Santa Cruz, Santa Cruz, CA).

Techniques: Infection, Control, shRNA, Staining