Journal: Communications Biology

Article Title: αvβ3 Integrin induces partial EMT independent of TGF-β signaling

doi: 10.1038/s42003-021-02003-6

Figure Lengend Snippet: a Representative immunohistochemistry images of β3 integrin expression in human normal lung and lung adenocarcinoma tumor tissues. Scale bar, 50 µm. b β3 integrin expression in human normal lung tissues ( n = 52) versus lung adenocarcinoma tumor tissues ( n = 112). The staining intensity was graded as follows: − (negative), + (weak), ++ (moderate), and +++ (strong). The source data are provided in Supplementary Data . c Kaplan-Meier analysis of the correlation between β3 integrin expression and overall survival for lung adenocarcinoma patients ( n = 720) using the database Kaplan-Meier Plotter. P values were calculated using a log-rank test.

Article Snippet: The fixed cells were stained with mouse monoclonal antibodies against αvβ3 integrin (clone LM609, #MAB1976, Merck Millipore, 1:100) and E-cadherin (clone HECD1, #M106, Takara, 1:2000), a goat polyclonal antibody against vimentin (clone C-20, #sc-7557, Santa Cruz Biotechnology, 1:100), a rat monoclonal antibody against CD31 (clone MEC13.3, #553370, BD Transduction Laboratories, 1:100), or a rabbit monoclonal antibody against Smad2/3 (clone D7G7, #8685, Cell Signaling Technology, 1:100) for 1 h at room temperature.

Techniques: Immunohistochemistry, Expressing, Staining

Journal: Communications Biology

Article Title: αvβ3 Integrin induces partial EMT independent of TGF-β signaling

doi: 10.1038/s42003-021-02003-6

Figure Lengend Snippet: a Correlation analysis between β3 integrin and the indicated EMT-related gene expression levels in human lung adenocarcinoma tissues by RNA sequencing from TCGA dataset. Spearman’s rank correlation and P value are shown ( n = 517 patients). b FACS analysis of the cell surface expression of αvβ3 integrin in control and β3 integrin expressing A549 cells. c Cell morphology of control and β3-A549 cells. Scale bar, 50 µm. d F-actin (red) and αvβ3 integrin (green) staining of control and β3-A549 cells. Images are shown with nuclear stain (blue). Scale bar, 25 µm. e Relative mRNA expression levels (qRT-PCR) of EMT markers in control and β3-A549 cells. Two-tailed unpaired Student’s t -test, mean ± s.e.m., n = 3 independent experiments conducted in triplicate. The source data are provided in Supplementary Data . f Western blotting of EMT markers in control and β3-A549 cells or control and β3-H358 cells. g Immunofluorescence-based single cell analysis using anti-E-cadherin and vimentin antibodies in control and β3-A549 cells. h Western blotting of EMT markers in H1975 cells expressing a control shRNA or two β3 integrin-specific shRNAs. i Cell morphology of H1975 cells expressing a control shRNA or two β3 integrin-specific shRNAs. Scale bar, 50 µm. Unprocessed blot images in ( f and h ) are shown in Supplementary Figs. .

Article Snippet: The fixed cells were stained with mouse monoclonal antibodies against αvβ3 integrin (clone LM609, #MAB1976, Merck Millipore, 1:100) and E-cadherin (clone HECD1, #M106, Takara, 1:2000), a goat polyclonal antibody against vimentin (clone C-20, #sc-7557, Santa Cruz Biotechnology, 1:100), a rat monoclonal antibody against CD31 (clone MEC13.3, #553370, BD Transduction Laboratories, 1:100), or a rabbit monoclonal antibody against Smad2/3 (clone D7G7, #8685, Cell Signaling Technology, 1:100) for 1 h at room temperature.

Techniques: Gene Expression, RNA Sequencing, Expressing, Control, Staining, Quantitative RT-PCR, Two Tailed Test, Western Blot, Immunofluorescence, Single-cell Analysis, shRNA

Journal: Communications Biology

Article Title: αvβ3 Integrin induces partial EMT independent of TGF-β signaling

doi: 10.1038/s42003-021-02003-6

Figure Lengend Snippet: a Representative images of migrated control and β3-A549 cells or control and β3-H460 cells in Boyden chamber assays. Scale bar, 100 µm. b , c Effect of αvβ3 integrin expression on the cell migration ( b ) and invasion ( c ) of A549, H358, and H460 cells. Two-tailed unpaired Student’s t -test, mean ± s.e.m. of three independent assays conducted in triplicate. d , e Effect of anti-αvβ3 integrin antibody ( d ) and β3 integrin-specific shRNA knockdown ( e ) on the cell migration of A549 ( d ) and H1975 ( e ) cells, respectively. One-way ANOVA and Bonferroni post-hoc test, mean ± s.e.m. of three independent assays conducted in triplicate. f Cell proliferation assays of control and β3-A549, or control and β3-H358 cells. Two-tailed unpaired Student’s t -test, mean ± s.e.m. of three independent assays conducted in quadruplicate. g Tumor growth of control and β3-A549 cells in nude mice. Two-tailed unpaired Student’s t -test, mean ± s.e.m., n = 5 mice per group. h Representative picture from the tumors of control and β3-A549 cells in nude mice at 4 weeks. Scale bar, 1 cm. i Primary tumor mass of control and β3-A549 cells in nude mice at 4 weeks. Two-tailed unpaired Student’s t -test, mean ± s.e.m., n = 5 mice per group. j , k Representative pictures from H&E-stained lungs ( j ), and quantification of lung colonies ( k ) at 7 weeks after tail vein injection of control and β3-A549 cells in nude mice. Arrowheads indicate tumor colonies. Scale bar, 1 mm. Two-tailed unpaired Student’s t -test, mean ± s.e.m., n = 6 mice per group. The source data are provided in Supplementary Data .

Article Snippet: The fixed cells were stained with mouse monoclonal antibodies against αvβ3 integrin (clone LM609, #MAB1976, Merck Millipore, 1:100) and E-cadherin (clone HECD1, #M106, Takara, 1:2000), a goat polyclonal antibody against vimentin (clone C-20, #sc-7557, Santa Cruz Biotechnology, 1:100), a rat monoclonal antibody against CD31 (clone MEC13.3, #553370, BD Transduction Laboratories, 1:100), or a rabbit monoclonal antibody against Smad2/3 (clone D7G7, #8685, Cell Signaling Technology, 1:100) for 1 h at room temperature.

Techniques: Control, Expressing, Migration, Two Tailed Test, shRNA, Knockdown, Staining, Injection

Journal: Communications Biology

Article Title: αvβ3 Integrin induces partial EMT independent of TGF-β signaling

doi: 10.1038/s42003-021-02003-6

Figure Lengend Snippet: a Western blotting of β3 integrin in control, β3, D119A β3, and ∆cyto β3 integrin-expressing A549 cells. b Cell adhesion of control, β3, D119A β3, and ∆cyto β3 integrin-expressing A549 cells to fibronectin, and the inhibitory effect of αvβ3 integrin antibody on the cell adhesion. c Phosphorylation of FAK in control, β3, D119A β3, and ∆cyto β3-expressing A549 cells. d Cell morphology of β3, D119A β3, and ∆cyto β3 integrin-expressing A549 cells. Scale bar, 50 µm. e Western blotting of EMT markers in control, β3, D119A β3, and ∆cyto β3 integrin-expressing A549 cells. f , g Cell migration ( f ) and invasion ( g ) of the indicated A549 cells. One-way ANOVA and Bonferroni post-hoc test, mean ± s.e.m. of three independent assays conducted in triplicate. The source data are provided in Supplementary Data . Unprocessed blot images in ( a , c and e ) are shown in Supplementary Fig. .

Article Snippet: The fixed cells were stained with mouse monoclonal antibodies against αvβ3 integrin (clone LM609, #MAB1976, Merck Millipore, 1:100) and E-cadherin (clone HECD1, #M106, Takara, 1:2000), a goat polyclonal antibody against vimentin (clone C-20, #sc-7557, Santa Cruz Biotechnology, 1:100), a rat monoclonal antibody against CD31 (clone MEC13.3, #553370, BD Transduction Laboratories, 1:100), or a rabbit monoclonal antibody against Smad2/3 (clone D7G7, #8685, Cell Signaling Technology, 1:100) for 1 h at room temperature.

Techniques: Western Blot, Control, Expressing, Phospho-proteomics, Migration

Journal: Communications Biology

Article Title: αvβ3 Integrin induces partial EMT independent of TGF-β signaling

doi: 10.1038/s42003-021-02003-6

Figure Lengend Snippet: a Cell morphology of control A549 cells, β3-A549 cells, and control and β3-A549 cells stably expressing the miR-200 family (β3-miR200-A549 cells) with or without 2 ng/ml TGF-β1 treatment for 24 h. Scale bar, 100 µm. b Relative mRNA expression levels (qRT-PCR) of EMT markers in β3-A549 and β3-miR200-A549 cells. One-way ANOVA and Bonferroni post-hoc test, mean ± s.e.m. of three independent assays conducted in triplicate. * P = 4.2 × 10 −11 , ** P = 7.6 × 10 −10 , *** P = 2.1 × 10 −8 . c Western blotting of EMT markers in β3-A549 and β3-miR200-A549 cells with or without 2 ng/ml TGF-β1 treatment for 24 h. d Effect of the miR-200 family expression on the cell migration of β3-A549 cells. Two-tailed unpaired Student’s t -test, mean ± s.e.m. of three independent assays conducted in triplicate. e Western blotting analysis of the intracellular expression levels of αv and β3 integrins in control, β3- and β3-miR200-A549 cells. f FACS analysis of the cell surface expression of αvβ3 integrin in control, β3-, and β3-miR200-A549 cells. The source data are provided in Supplementary Data . Unprocessed blot images in ( c and e ) are shown in Supplementary Fig. .

Article Snippet: The fixed cells were stained with mouse monoclonal antibodies against αvβ3 integrin (clone LM609, #MAB1976, Merck Millipore, 1:100) and E-cadherin (clone HECD1, #M106, Takara, 1:2000), a goat polyclonal antibody against vimentin (clone C-20, #sc-7557, Santa Cruz Biotechnology, 1:100), a rat monoclonal antibody against CD31 (clone MEC13.3, #553370, BD Transduction Laboratories, 1:100), or a rabbit monoclonal antibody against Smad2/3 (clone D7G7, #8685, Cell Signaling Technology, 1:100) for 1 h at room temperature.

Techniques: Control, Stable Transfection, Expressing, Quantitative RT-PCR, Western Blot, Migration, Two Tailed Test

Journal: Bioscience Reports

Article Title: Reversal of EGFR inhibitors’ resistance by co-delivering EGFR and integrin αvβ3 inhibitors with nanoparticles in non-small cell lung cancer

doi: 10.1042/BSR20181259

Figure Lengend Snippet: ( A ) The expression of integrin αvβ3 + tumor cells in EGFR inhibitor drugs-resistant (EIR) and -sensitive tumor (EIS) tissues obtained from NSCLC patients via using immunohistochemistry (IHC) (bar, 50 μm). ( B ) The percentage of integrin αvβ3 + tumor cells in EGFR inhibitor drugs-resistant and -sensitive tumor tissues obtained from NSCLC patients by using flow cytometry. ( C ) Cell viability of total, integrin αvβ3 − and αvβ3 + separated from A549, NCI-H1975 (H1975) and Lewis cells after treated with Erlo (10 μM) for 48 h. ( D ) Cell viability of total, integrin αvβ3 − and αvβ3 + from A549, NCI-H1975 and Lewis cells after treating with Lapa (10 μM) for 48 h. ( E ) Cell viability of total, integrin αvβ3 − and αvβ3 + from A549, NCI-H1975 and Lewis cells after treated with Gefi (3 μM for A549 and 10 μM for NCI-H1975/Lewis, respectively) for 48 h. ( F ) Cell viability of total, integrin αvβ3 − and αvβ3 + from A549, NCI-H1975 and Lewis cells after treated with doxorubicin (10 nM) for 48 h. ( G ) Cell viability of total, integrin αvβ3 − and αvβ3 + from A549, NCI-H1975 and Lewis cells after treated with PTX (30 nM for A549/NCI-H1975 cells and 60 nM for Lewis cells, respectively) for 48 h. ( H ) Cell viability of total, integrin αvβ3 − and αvβ3 + from A549, NCI-H1975 and Lewis cells after treated with GEM (1 μM for A549/NCI-H1975 cells and 3 μM for Lewis cells, respectively) for 48 h. ( I ) Tumor volume of nude mice bearing total, integrin αvβ3 − and αvβ3 + separated from A549 cells treated with PBS or Erlo (50 mg/kg) were measured ( n =8) (up). Survival time of nude mice bearing total, integrin αvβ3 − and αvβ3 + from A549 cells treated with PBS or Erlo (50 mg/kg) in each group were recorded ( n =8) (down). ( J ) Tumor volume of nude mice bearing total, integrin αvβ3 − and αvβ3 + from A549 cells treated with PBS or Lapa (100 mg/kg) were measured ( n =8) (left). Survival time of nude mice model bearing total, integrin αvβ3 − and αvβ3 + from A549 cells treated with PBS or Lapa (100 mg/kg) in each group were recorded ( n =8) (right). ( K ) Tumor volume of nude mice bearing total, integrin αvβ3 − and αvβ3 + separated from A549 cells treated with PBS or Gefi (200 mg/kg) were measured ( n =8) (left). Survival time of nude mice model bearing total, integrin αvβ3 − and αvβ3 + from A549 cells treated with PBS or Gefi (200 mg/kg) in each group were recorded ( n =8) (right). * P <0.05; ** P <0.01; *** P <0.001; ns, no significant difference.

Article Snippet: Samples were solubilized with an equal volume of loading buffer (125 mM Tris/HCl, pH 6.8, 4% sodium dodecyl sulfate, 20% glycerol, 0.05% Bromophenol Blue, 5% β-mercaptoethanol) and were boiled for 10 min, then samples were separated by SDS/PAGE, followed by transferring to PVDF membranes and detecting by immunoblotting with primary antibodies against human integrin αvβ3 (Abcam, ab119992, 1:500, Cambridge, U.K.), Galectin-3 (CST, 87985, 1:1000, Boston, U.S.A.), KRAS (Abcam, ab180772, 1:200, Cambridge, U.K.), RalB (CST, 3523, 1:1000, Boston, U.S.A.), TBK1 (CST, 3504, 1:1000, Boston, U.S.A.), p-TBK1 (CST, 5483, 1:1000, Boston, U.S.A.), respectively, at 4°C overnight.

Techniques: Expressing, Immunohistochemistry, Flow Cytometry

Journal: Bioscience Reports

Article Title: Reversal of EGFR inhibitors’ resistance by co-delivering EGFR and integrin αvβ3 inhibitors with nanoparticles in non-small cell lung cancer

doi: 10.1042/BSR20181259

Figure Lengend Snippet: ( A ) The percent of integrin αvβ3 + cells in A549 received PBS, Erlo (10 μM), Lapa (10 μM) or Gefi (3 μM) treatment was detected by using flow cytometry. ( B ) The percent of integrin αvβ3 + cells in NCI-H1975 received PBS, Erlo (10 μM), Lapa (10 μM) and Gefi (10 μM) treatment was detected by using flow cytometry. ( C ) The percent of integrin αvβ3 + cells in Lewis received PBS, Erlo (10 μM), Lapa (10 μM) and Gefi (10 μM) treatment was detected by using flow cytometry. ( D ) A549, NCI-H1975 and Lewis cells were pre-treated with Erlo (10 μM) for 48 h. Then the living cells were retreated by Erlo (10 μM) for another 48 h, followed by cell viability detection ( n =3). ( E ) A549, NCI-H1975 and Lewis cells were pre-treated with Lapa (10 μM) for 48 h. Then the living cells were retreated by Lapa (10 μM) for another 48 h, followed by cell viability detection ( n =3). ( F ) A549, NCI-H1975 and Lewis cells were pre-treated with Gefi (3/10/10 μM) for 48 h. Then the living cells were retreated by Gefi (3/10/10 μM) for another 48 h, followed by cell viability detection ( n =3). ( G ) The nude mice bearing A549 cells were treated with PBS, Erlo (50 mg/kg), Lapa (100 mg/kg) or Gefi (50 mg/kg) for 3 days, then the percent of integrin αvβ3 + cells in tumor tissue were detected by using flow cytometry ( n =8). ( H ) The nude mice bearing A549 cells were treated with PBS or Erlo (50 mg/kg) for 3 days, then the tumor tissues were excised and treated with PBS, Erlo (10 μM), Lapa (10 μM) or Gefi (10 μM) respectively for 48 h in vitro . Then the apoptotic rate was detected by using flow cytometry ( n =5).

Article Snippet: Samples were solubilized with an equal volume of loading buffer (125 mM Tris/HCl, pH 6.8, 4% sodium dodecyl sulfate, 20% glycerol, 0.05% Bromophenol Blue, 5% β-mercaptoethanol) and were boiled for 10 min, then samples were separated by SDS/PAGE, followed by transferring to PVDF membranes and detecting by immunoblotting with primary antibodies against human integrin αvβ3 (Abcam, ab119992, 1:500, Cambridge, U.K.), Galectin-3 (CST, 87985, 1:1000, Boston, U.S.A.), KRAS (Abcam, ab180772, 1:200, Cambridge, U.K.), RalB (CST, 3523, 1:1000, Boston, U.S.A.), TBK1 (CST, 3504, 1:1000, Boston, U.S.A.), p-TBK1 (CST, 5483, 1:1000, Boston, U.S.A.), respectively, at 4°C overnight.

Techniques: Flow Cytometry, In Vitro

Journal: Bioscience Reports

Article Title: Reversal of EGFR inhibitors’ resistance by co-delivering EGFR and integrin αvβ3 inhibitors with nanoparticles in non-small cell lung cancer

doi: 10.1042/BSR20181259

Figure Lengend Snippet: ( A ) The expressions of integrin αvβ3, Glatin-3, KRAS, RalB, p-TBK1, TBK1 and actin in integrin αvβ3 − and αvβ3 + A549 cells were detected by using Western blot. ( B ) The distribution of NF-κB in integrin αvβ3 − and αvβ3 + A549 cells were examined by using confocal (bar, 50 μM). ( C ) Cell viability of A549, NCI-H1975 and Lewis cells treated with Erlo (10 μM) with or without Cilen (5 μM) for 48 h. ( D ) Cell viability of A549, NCI-H1975 and Lewis cells treated Lapa (10 μM) with or without Cilen (5 μM) for 48 h. ( E ) Cell viability of A549, NCI-H1975 and Lewis cells treated Gefi (3/10/10 μM) with or without Cilen (5 μM) for 48 h. ( F ) Tumor volume of nude mice bearing integrin αvβ3 + A549 cells treated with PBS, Erlo (50 mg/kg) or Erlo (50 mg/kg) combining with Cilen (200 mg/kg) were measured ( n =8) (left). Survival time of nude mice bearing αvβ3 + A549 cells treated with PBS, Erlo (50 mg/kg) with Cilen (200 mg/kg) were recorded ( n =8) (right). ( G ) The body weight of C57 mice receiving PBS, Erlo (50 mg/kg) and Erlo (50 mg/kg) + Cilen (200 mg/kg) ( n =6) treatment was measured. ( H ) The concentrations of CRE were detected in C57 mice receiving PBS, Erlo (50 mg/kg) and Erlo (50 mg/kg) + Cilen (200 mg/kg). ( I ) The levels of GPT were detected in C57 mice treated with PBS, Erlo (50 mg/kg) and Erlo (50 mg/kg) + Cilen (200 mg/kg). ( J ) The levels of GOT were detected in in C57 mice received PBS, Erlo (50 mg/kg) and Erlo (50 mg/kg) + Cilen (200 mg/kg). ** P <0.01; *** P <0.001, ns, not statistically significant.

Article Snippet: Samples were solubilized with an equal volume of loading buffer (125 mM Tris/HCl, pH 6.8, 4% sodium dodecyl sulfate, 20% glycerol, 0.05% Bromophenol Blue, 5% β-mercaptoethanol) and were boiled for 10 min, then samples were separated by SDS/PAGE, followed by transferring to PVDF membranes and detecting by immunoblotting with primary antibodies against human integrin αvβ3 (Abcam, ab119992, 1:500, Cambridge, U.K.), Galectin-3 (CST, 87985, 1:1000, Boston, U.S.A.), KRAS (Abcam, ab180772, 1:200, Cambridge, U.K.), RalB (CST, 3523, 1:1000, Boston, U.S.A.), TBK1 (CST, 3504, 1:1000, Boston, U.S.A.), p-TBK1 (CST, 5483, 1:1000, Boston, U.S.A.), respectively, at 4°C overnight.

Techniques: Western Blot

Journal: Bioscience Reports

Article Title: Reversal of EGFR inhibitors’ resistance by co-delivering EGFR and integrin αvβ3 inhibitors with nanoparticles in non-small cell lung cancer

doi: 10.1042/BSR20181259

Figure Lengend Snippet: ( A ) Tumor volume of nude mice bearing integrin αvβ3 + A549 cells treated with PBS, Erlo (50 mg/kg) combined with Cilen (200 mg/kg) or MPEG-PLA encapsulated Erlo (50 mg/kg) and Cilen (200 mg/kg) were measured ( n =8) (upper). Survival time of nude mice bearing integrin αvβ3 + A549 cells treated with PBS, Erlo (50 mg/kg) combined with Cilen (200 mg/kg) or MPEG-PLA encapsulated Erlo (50 mg/kg) and Cilen (200 mg/kg) in each group were recorded ( n =8) (down). ( B ) Tumor volume of nude mice bearing integrin αvβ3 + A549 cells treated with PBS, Lapa (100 mg/kg) combined with Cilen (200 mg/kg) or MPEG-PLA encapsulated Lapa (100 mg/kg) and Cilen (200 mg/kg) were measured ( n =8) (upper). Survival time of nude mice bearing integrin αvβ3 + A549 cells treated with PBS, Lapa (100 mg/kg) combined with Cilen (200 mg/kg) or MPEG-PLA encapsulated Lapa (100 mg/kg) and Cilen (200 mg/kg) in each group were recorded ( n =8) (down). ( C ) Tumor volume of nude mice bearing integrin αvβ3 + A549 cells treated with PBS, Gefi (50 mg/kg) combining with Cilen (200 mg/kg) or MPEG-PLA encapsulated Gefi (50 mg/kg) and Cilen (200 mg/kg) were measured ( n =8) (upper). Survival time of nude mice bearing integrin αvβ3 + A549 cells treated with PBS, Gefi (50 mg/kg) combined with Cilen (200 mg/kg) or MPEG-PLA encapsulated Gefi (50 mg/kg) and Cilen (200 mg/kg) in each group were recorded ( n =8) (down). ( D ) The body weight of C57 mice was measured in PBS, Erlo (50 mg/kg) + Cilen (200 mg/kg) or MPEG-PLA encapsulated Erlo (50 mg/kg) + Cilen (200 mg/kg) groups ( n =6). ( E ) The concentrations of CRE were detected in C57 mice received PBS, Erlo (50 mg/kg) + Cilen (200 mg/kg) or MPEG-PLA encapsulating Erlo (50 mg/kg) + Cilen (200 mg/kg). ( F ) The levels of GPT were detected in C57 mice treated with PBS, Erlo (50 mg/kg) + Cilen (200 mg/kg) or MPEG-PLA encapsulating Erlo (50 mg/kg) + Cilen (200 mg/kg). ( G ) The levels of GOT were detected in C57 mice receiving PBS, Erlo (50 mg/kg) + Cilen (200 mg/kg) or MPEG-PLA encapsulating Erlo (50 mg/kg) + Cilen (200 mg/kg). ( H ) The schematic diagram of EGFR inhibitor resistance induced by integrin αvβ3 in NSCLC. * P <0.05; ** P <0.01; *** P <0.001; ns, no significant difference.

Article Snippet: Samples were solubilized with an equal volume of loading buffer (125 mM Tris/HCl, pH 6.8, 4% sodium dodecyl sulfate, 20% glycerol, 0.05% Bromophenol Blue, 5% β-mercaptoethanol) and were boiled for 10 min, then samples were separated by SDS/PAGE, followed by transferring to PVDF membranes and detecting by immunoblotting with primary antibodies against human integrin αvβ3 (Abcam, ab119992, 1:500, Cambridge, U.K.), Galectin-3 (CST, 87985, 1:1000, Boston, U.S.A.), KRAS (Abcam, ab180772, 1:200, Cambridge, U.K.), RalB (CST, 3523, 1:1000, Boston, U.S.A.), TBK1 (CST, 3504, 1:1000, Boston, U.S.A.), p-TBK1 (CST, 5483, 1:1000, Boston, U.S.A.), respectively, at 4°C overnight.

Techniques:

Journal: Translational research : the journal of laboratory and clinical medicine

Article Title: Emerging strategies to disrupt the central TGF-β axis in kidney fibrosis

doi: 10.1016/j.trsl.2019.04.003

Figure Lengend Snippet: Selected antifibrotic agents in clinical development that modulate TGF-β activation, expression, or signaling

Article Snippet: An antibody targeting αvβ6 (BG0001, Biogen) is in a phase II trial to treat pulmonary fibrosis, while a blocking antibody against αvβ3 (VPI-2690B, Vascular Pharmaceuticals) is currently being tested in a phase 2 trial of diabetic nephropathy.

Techniques: Activation Assay, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Peptide-Guided Photodynamic Therapy via Integrin αvβ6 in Pancreatic Cancer

doi: 10.3390/ijms27041838

Figure Lengend Snippet: ( a ) Expression of αvβ6 and αvβ3 integrins in pancreatic cancer cell lines. ( a ) Normalized fluorescence measurements showing integrin expression levels in BxPc3, MiaPaca2, and Panc1 cells. BxPc3 cells display significantly higher αvβ6 expression compared with MiaPaca2 and Panc1, whereas αvβ3 expression is detectable at lower levels with a different expression profile across cell lines. Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by post hoc multiple comparison testing; differences were considered statistically significant at *** p < 0.001 (* p < 0.05 where indicated). ( b ) Western blot results showing β6 integrin expression levels normalized to protein concentration (pg β/µg protein) across different PDAC cell lines. BxPc3 expresses high levels of β6 (1411 pg/µg), MiaPaca2 and Panc1 expresses show no detectable levels of this integrin.

Article Snippet: Primary antibodies included Anti-Integrin αvβ3 (Integrin β3 (D7X3P) XP ® Rabbit mAb, Cell Signaling) at a 1:500 dilution and Anti-Integrin αvβ6 (clone EM05201, ZooMAb ® Rabbit Monoclonal ZRB1104) at a 1:1000 dilution, incubated overnight at 4 °C.

Techniques: Expressing, Fluorescence, Comparison, Western Blot, Protein Concentration