Journal: American Journal of Translational Research

Article Title: Lyn kinase enhanced hepatic fibrosis by modulating the activation of hepatic stellate cells

doi:

Figure Lengend Snippet: Increased activation of HSCs in Lyn TG mice upon CCl4 treatment. A: Representative immunofluorescence photomicrographs of HSC activation marker (Αvβ3, α-SMA and desmin) expression in liver tissues from WT mice and Lyn TG mice after administration of CCl4 (×200 magnification). B: The fluorescence intensity of desmin in 8 random fields. C: The fluorescence intensity of α-SMA in 8 random fields. D: The fluorescence intensity of Αvβ3 in 8 random fields. E: TGF-β1 levels in the liver tissues were determined by ELISA. All of the data are presented as the mean ± s.d. One-way ANOVA (Tukey-Kramer post-test) was used for Statistical analysis.

Article Snippet: After blocking with 1% BSA (Sigma-Aldrich) containing 0.05% Tween 20 for 1 hour, the specimens were then incubated with anti-Lyn antibody (Santa Cruz, CA, sc-15, AB_2281450), anti-integrin αvβ3 antibody (Bioss, China, bs-1310R), anti-α-SMA antibody (Bioss, China, bs-0189R), anti-desmin antibody (Bioss, China, bs-1026R), and anti-COL1α1 antibody (Bioss, China, bs-10423R).

Techniques: Activation Assay, Immunofluorescence, Marker, Expressing, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: American Journal of Translational Research

Article Title: Lyn kinase enhanced hepatic fibrosis by modulating the activation of hepatic stellate cells

doi:

Figure Lengend Snippet: Lyn kinase increased the activation of HSCs in vitro. Immunohistochemical staining for HSC activation marker (Αvβ3, α-SMA and COL1α1) expression in TGF-β1-induced LX-2 and Lyn+/+ LX-2. A: Representative photomicrographs and fluorescence intensity of α-SMA in cells (×400 magnification). B: Representative photomicrographs and fluorescence intensity of Αvβ3 in cells (×400 magnification). C: Representative photomicrographs and fluorescence intensity of COL1α1 in cells (×400 magnification). All of the data are presented as the mean ± s.d. One-way ANOVA (Tukey-Kramer post-test) was used for Statistical analysis. LX-2 cell lines transfected with lentiviral vector expressing Lyn were described as Lyn+/+ cells. LX-2 cell lines transfected with control lentiviral vector were described as WT cells.

Article Snippet: After blocking with 1% BSA (Sigma-Aldrich) containing 0.05% Tween 20 for 1 hour, the specimens were then incubated with anti-Lyn antibody (Santa Cruz, CA, sc-15, AB_2281450), anti-integrin αvβ3 antibody (Bioss, China, bs-1310R), anti-α-SMA antibody (Bioss, China, bs-0189R), anti-desmin antibody (Bioss, China, bs-1026R), and anti-COL1α1 antibody (Bioss, China, bs-10423R).

Techniques: Activation Assay, In Vitro, Immunohistochemical staining, Staining, Marker, Expressing, Fluorescence, Transfection, Plasmid Preparation

Journal: American Journal of Translational Research

Article Title: Lyn kinase enhanced hepatic fibrosis by modulating the activation of hepatic stellate cells

doi:

Figure Lengend Snippet: The Src-specific inhibitor PP2 suppressed HSC activation. A: Immunohistochemical staining for HSC activation marker (Αvβ3) expression in TGF-β1-induced LX-2 and Lyn+/+ LX-2 in the presence of PP2 (×400 magnification). The fluorescence intensity of Αvβ3 in cells was measured. B: Apoptotic cells of LX-2 were detected by flow cytometry. All of the data are presented as the mean ± s.d. One-way ANOVA (Tukey-Kramer post-test) was used for Statistical analysis.

Article Snippet: After blocking with 1% BSA (Sigma-Aldrich) containing 0.05% Tween 20 for 1 hour, the specimens were then incubated with anti-Lyn antibody (Santa Cruz, CA, sc-15, AB_2281450), anti-integrin αvβ3 antibody (Bioss, China, bs-1310R), anti-α-SMA antibody (Bioss, China, bs-0189R), anti-desmin antibody (Bioss, China, bs-1026R), and anti-COL1α1 antibody (Bioss, China, bs-10423R).

Techniques: Activation Assay, Immunohistochemical staining, Staining, Marker, Expressing, Fluorescence, Flow Cytometry

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:

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:

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:

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:

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

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:

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

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

Journal: Journal of Virology

Article Title: Pathogenic Old World Hantaviruses Infect Renal Glomerular and Tubular Cells and Induce Disassembling of Cell-to-Cell Contacts ^▿

doi: 10.1128/JVI.00568-11

Figure Lengend Snippet: Expression of marker proteins and the hantaviral receptor integrin αVβ3 on renal cell types. (A) Human primary cells HREpC, HRGEnC, and human podocytes were stained with antibodies against marker proteins for renal cell types and with anti-integrin αVβ3 antibody. (B) Lysates of renal cell types were analyzed for the expression of integrin β3 by Western blot analysis. (C) Flow cytometric analysis of cell surface protein expression of integrin αVβ3 and the endothelial marker CD31.

Article Snippet: To confirm the specificity of anti-integrin α V β 3 antibody LM609, fixed cells were incubated with anti-integrin α V β 3 antibody that was pretreated with recombinant human integrin α V β 3 (R&D Systems, Wiesbaden-Nordenstadt, Germany).

Techniques: Expressing, Marker, Staining, Western Blot

Journal: Journal of Virology

Article Title: Pathogenic Old World Hantaviruses Infect Renal Glomerular and Tubular Cells and Induce Disassembling of Cell-to-Cell Contacts ^▿

doi: 10.1128/JVI.00568-11

Figure Lengend Snippet: Characterization of human renal cells and specificity of the anti-integrin antibodies. (A) HREpC, HRGEnC, and podocytes were analyzed for the presence or absence of the epithelial marker cytokeratin 18, the endothelial marker CD31, and the podocyte-specific protein synaptopodin, along with the specific isotype control antibodies. (B) Analysis of the specificity the of anti-integrin αVβ3 antibody LM609. Fixed renal cells were incubated with isotype control antibody, with anti-integrin αVβ3 LM609 or with anti-integrin αVβ3 antibody LM609 that was preincubated with recombinant human integrin αVβ3. (C) Specificity of the anti-integrin β3 antibody was confirmed by the absence of an integrin-specific band in K562 lysate lacking integrin expression (30).

Article Snippet: To confirm the specificity of anti-integrin α V β 3 antibody LM609, fixed cells were incubated with anti-integrin α V β 3 antibody that was pretreated with recombinant human integrin α V β 3 (R&D Systems, Wiesbaden-Nordenstadt, Germany).

Techniques: Marker, Incubation, Recombinant, Expressing

Journal: Journal of Virology

Article Title: Pathogenic Old World Hantaviruses Infect Renal Glomerular and Tubular Cells and Induce Disassembling of Cell-to-Cell Contacts ^▿

doi: 10.1128/JVI.00568-11

Figure Lengend Snippet: Expression of hantaviral receptor integrin αVβ3 in tubules and glomeruli of human kidney. Human renal cryosections were fixed and stained with antibodies against the hantaviral receptor integrin αVβ3 (red) and the marker for endothelial cells CD31 (green, upper row) or the marker for podocytes, synaptopodin (green, lower row). Tubules are marked with asterisks, and peritubular capillaries are marked with arrowheads.

Article Snippet: To confirm the specificity of anti-integrin α V β 3 antibody LM609, fixed cells were incubated with anti-integrin α V β 3 antibody that was pretreated with recombinant human integrin α V β 3 (R&D Systems, Wiesbaden-Nordenstadt, Germany).

Techniques: Expressing, Staining, Marker

Journal: Advanced Science

Article Title: Self‐Triggered Apoptosis Enzyme Prodrug Therapy (STAEPT): Enhancing Targeted Therapies via Recurrent Bystander Killing Effect by Exploiting Caspase‐Cleavable Linker

doi: 10.1002/advs.201800368

Figure Lengend Snippet: Enhanced cellular uptake and cytotoxic activity of RGDEVD‐DOX in integrin αvβ3 expressing cells. a) Representative confocal images (left) and quantitative analysis (right) of HDMEC and U‐87 MG cells exposed to fluorescent‐labeled RDEVD and RGDEVD peptides. b) Representative confocal images (left) and quantitative analysis (right) of scrambled control and integrin αv (ITGAV) siRNA‐transfected HDMECs and U‐87 MG cells exposed to fluorescent‐labeled RGDEVD peptide. Green and blue indicate the fluorescent‐labeled peptides and cell nuclei, respectively. Scale bar, 50 µm. c) Flow cytometry analysis of isotype control (top), scrambled control‐transfected (lower left), and ITGAV siRNA‐transfected (lower right) U87 MG cells incubated with fluorescent‐labeled RGDEVD and stained with an antibody against integrin αvβ3. d) Representative confocal images (left) and quantitative analysis (right) of U‐87 MG and HT‐29 cells treated with RDEVD‐DOX and RGDEVD‐DOX. Red and blue indicate the intrinsic red fluorescence of doxorubicin and cell nuclei, respectively. Scale bar, 50 µm. e) Concentration‐dependent cytotoxicity of RDEVD‐DOX and RGDEVD‐DOX on U‐87 MG (left) and HT‐29 (right) determined by MTT assay ( n = 4). f) Doxorubicin release from RGDEVD‐DOX when incubated in PBS (pH 7.4) containing (or not containing) carboxylesterase ( n = 3). Data are mean ± s.d. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The cells were incubated with Alexa Fluor 488‐conjugated integrin αvβ3 antibody (1:100; R&D Systems, Minneapolis, MN; Cat. No. FAB3050G) for an hour at 4 °C, washed, and suspended in PBS containing 0.5% BSA.

Techniques: Activity Assay, Expressing, Labeling, Control, Transfection, Flow Cytometry, Incubation, Staining, Fluorescence, Concentration Assay, MTT Assay

Journal: Biology

Article Title: Engineering a Dual Specificity γδ T-Cell Receptor for Cancer Immunotherapy

doi: 10.3390/biology13030196

Figure Lengend Snippet: Evaluation of αvβ6-dependent activation of chimeric G115 γδ TCRs. ( A ) Analysis of integrin expression on A375 puro and A375-β6 cells. Red–integrin; blue–isotype control. Data are representative of 3 independent replicates. Co-cultures were performed between A375 puro ( B ) and A375-β6 ( C ) tumour cells and untransduced (untrans) or transduced T-cell populations at an effector-to-target ratio of 1:1 for 72 h. Residual tumour cell viability was determined using an MTT assay (mean ± SEM of indicated replicates). Statistical analysis was performed using one-way ANOVA; **** p < 0.0001. ( D ) Supernatants collected from co-cultures described in ( B , C ) were analysed for IFN-γ by ELISA (mean ± SEM; n = 21–45 from 4–6 independent donors). Statistical analysis was performed using one-way ANOVA; * p < 0.05; ** p < 0.01.

Article Snippet: Integrin expression was detected by flow cytometry using the following antibodies: αvβ3 integrin (FAB3050A, R&D Systems, Minneapolis, MN, USA), αvβ5 integrin (FAB2528A, R&D systems), αvβ6 integrin (FAB4155A, R&D Systems) and αvβ8 integrin (FAB4775A, R&D systems).

Techniques: Activation Assay, Expressing, Control, MTT Assay, Enzyme-linked Immunosorbent Assay

Journal: Biology

Article Title: Engineering a Dual Specificity γδ T-Cell Receptor for Cancer Immunotherapy

doi: 10.3390/biology13030196

Figure Lengend Snippet: Evaluation of PAg-dependent activation of chimeric G115 γδ TCRs by ffLuc-expressing K562 cells. ( A ) Analysis of αvβ6 integrin expression on ffLuc + K562 cells. Red–integrin; blue–isotype control. Data are representative of 3 independent replicates. ( B ) Firefly luciferase-expressing K562 cells were pre-incubated with the indicated Zol concentration for 24 h prior to the establishment of co-cultures with untrans(duced) or transduced T-cell populations at an effector to target ratio 1:1 for 72 h. Data show mean ± SEM of residual K562 viability (n = 6–11 from 4 independent donors), as determined by luciferase assay. Statistical analysis was performed using two-way ANOVA; *** p < 0.001; **** p < 0.0001. ( C ) Supernatants collected from co-cultures described in B were analysed for IFN-γ by ELISA (mean ± SEM; n = 19 from 3 independent donors). Statistical analysis was performed using two-way ANOVA; *** p < 0.001; **** p < 0.0001.

Article Snippet: Integrin expression was detected by flow cytometry using the following antibodies: αvβ3 integrin (FAB3050A, R&D Systems, Minneapolis, MN, USA), αvβ5 integrin (FAB2528A, R&D systems), αvβ6 integrin (FAB4155A, R&D Systems) and αvβ8 integrin (FAB4775A, R&D systems).

Techniques: Activation Assay, Expressing, Control, Luciferase, Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Biology

Article Title: Engineering a Dual Specificity γδ T-Cell Receptor for Cancer Immunotherapy

doi: 10.3390/biology13030196

Figure Lengend Snippet: Evaluation of the anti-tumour activity of chimeric G115 γδ TCRs against BxPC3 pancreatic tumour cells. ( A ) Analysis of integrin expression on BxPC3 cells. Red–integrin; blue–isotype control. Data are representative of 3 independent replicates. Co-cultures were performed between BxPC3 tumour cells (No Zol; B ) or Zol-sensitised BxPC3 tumour cells (+Zol; C ) and untrans(duced) or transduced T-cell populations at an effector to target ratio of 1:1 for 72 h. Tumour cell viability was determined using an MTT assay (mean ± SEM of indicated replicates). Statistical analysis was performed using one-way ANOVA; **** p < 0.0001, * p < 0.05, N/S–not significant.

Article Snippet: Integrin expression was detected by flow cytometry using the following antibodies: αvβ3 integrin (FAB3050A, R&D Systems, Minneapolis, MN, USA), αvβ5 integrin (FAB2528A, R&D systems), αvβ6 integrin (FAB4155A, R&D Systems) and αvβ8 integrin (FAB4775A, R&D systems).

Techniques: Activity Assay, Expressing, Control, MTT Assay

Journal: Biology

Article Title: Engineering a Dual Specificity γδ T-Cell Receptor for Cancer Immunotherapy

doi: 10.3390/biology13030196

Figure Lengend Snippet: Evaluation of the anti-tumour activity of chimeric G115 γδ TCRs against Panc1 pancreatic tumour cells. ( A ) Analysis of integrin expression on Panc1 cells. Red–integrin; blue–isotype control. Data are representative of 3 independent replicates. Co-cultures were performed between Panc1 tumour cells (No Zol; B ) or Zol-sensitised Panc1 tumour cells (+ Zol; C ) and untransduced (untrans) or transduced T-cell populations at an effector to target ratio of 1:1 for 72 h. Tumour cell viability was determined using an MTT assay (mean ± SEM of indicated replicates). Statistical analysis was performed using one-way ANOVA; **** p < 0.0001, ** p < 0.01, N/S–not significant.

Article Snippet: Integrin expression was detected by flow cytometry using the following antibodies: αvβ3 integrin (FAB3050A, R&D Systems, Minneapolis, MN, USA), αvβ5 integrin (FAB2528A, R&D systems), αvβ6 integrin (FAB4155A, R&D Systems) and αvβ8 integrin (FAB4775A, R&D systems).

Techniques: Activity Assay, Expressing, Control, MTT Assay

Journal: iScience

Article Title: TGF-β1 promotes osteogenesis of mesenchymal stem cells via integrin mediated mechanical positive autoregulation

doi: 10.1016/j.isci.2024.110262

Figure Lengend Snippet:

Article Snippet: The primary antibodies used here were as follows: anti-Runx2 antibody (1:100; Abcam, USA), anti-integrin alpha V + beta 3 antibody (1: 200; GeneTex, USA), anti-paxillin antibody (1: 100; Abcam, USA), anti-phospho-FAK antibody (1: 100; Abcam, USA), anti-YAP antibody (1: 200; Cell Signaling Technology, USA) and anti-lamin A/C antibody (1: 200; Cell Signaling Technology, USA).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, RNA Extraction, Real-time Polymerase Chain Reaction, Software