mouse monoclonal integrin β3  (R&D Systems)

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: MV leads to upregulation of integrin β3 and PKM2 in pulmonary fibroblasts. Mice lung tissue were acquired at Day 7 after 2 h of MV. A , functional enrichment analysis with GO Biological Processes was performed using GOrilla with the differential genes upregulated and downregulated in pulmonary fibroblasts cluster. B , heatmaps are showing the upregulated and downregulated genes of integrin and glycolysis in pulmonary fibroblasts cluster. C , protein expression of integrin β3 and pyruvate kinase M2 (PKM2) in lung homogenates was assessed by immunohistochemistry and quantified by the average optical density value (AOD), n = 3 per group. D , protein expression of integrin β3 and PKM2 in lung homogenates was determined by western blot analysis. Relative densitometry of the protein bands of integrin β3 and PKM2 over β-actin is displayed in bar graphs, n = 3 in sham group and n = 9 in MV group. Data are expressed as means ± SEM. *p < 0.05, **p < 0.01, *** p < 0.001(t test).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Functional Assay, Expressing, Immunohistochemistry, Western Blot

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: MV leads to upregulation of integrin β3 and PKM2 in pulmonary fibroblasts. Mice lung tissue were acquired at Day 7 after 2 h of MV. A and B, lung tissues were stained with fluorophore-labeled antibodies against fibroblast marker fibronectin (FN) (Alexa Fluor 568, red), integrin β3(β3) (fluorescein isothiocyanate, green) and PKM2 (fluorescein isothiocyanate, green). 4',6-diamidino-2- phenylindole (DAPI) stain was used to detect nuclei (blue). Original magnification × 400. Scale bars correspond to 20 µm. C , flow cytometry was used to detect the expression of integrin β3 and pyruvate kinase M2 (PKM2) in lung cells and in fibronectin (FN) positive fibroblasts, n = 6 per group. Data are expressed as means ± SEM. *p < 0.05, **p < 0.01, *** p < 0.001(t test).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Staining, Labeling, Marker, Flow Cytometry, Expressing

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: Integrin β3 inhibition ameliorates PKM2-mediated aerobic glycolysis and MV-induced pulmonary fibrosis. A and B , both wild-type and integrin β3-deficient (β3 +/- ) mice were subjected to mechanical ventilation (MV) for 2 h. Lung tissues were harvested at day 7. A , lung fibrosis was quantified by Masson's trichrome staining, original magnification × 200, scale bars correspond to 100μm, n = 6, 5, 6, 6, respectively in each group. B , integrin β3, pyruvate kinase M2 (PKM2), lactic dehydrogenase (LDHA), collagen I α1 (COL1A1) and α smooth muscle actin (α-SMA) expression was quantified by western blot analysis, n = 3 per group. C and D , mice were treated intraperitoneally with integrin β3 inhibitor cilengitide 4 mg/kg once a day for 3 consecutive days before being subjected to MV for 2 h. Lung tissues were harvested 7 days after intubation. C , lung fibrosis was quantified by Masson's trichrome staining, original magnification × 200, scale bars correspond to 100 µm, n = 6 per group. D , integrin β3, PKM2, LDHA and COL1A1 expression was quantified by western blot analysis, n = 3 per group. Data are expressed as means ± SEM, *p < 0.05, **p < 0.01, *** p < 0.001 (ANOVA).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Inhibition, Staining, Expressing, Western Blot

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: Integrin β3 inhibition ameliorates PKM2-mediated aerobic glycolysis and MV-induced pulmonary fibrosis . A-D , both wild-type and integrin β3-deficient (β3 +/- ) mice were subjected to mechanical ventilation (MV) for 2 h. Lung tissues were harvested at day 7. A and B , lung tissues were stained with fluorophorelabeled antibodies against fibroblast marker fibronectin (FN) (red), pyruvate kinase M2 (PKM2) (green) and collagen I (green), 4',6-diamidino-2- phenylindole (DAPI) stain was used to detect nuclei (blue), original magnification × 400, scale bars correspond to 20 µm. C and D , lactate (Lac) and type I procollagen carboxyterminal peptide (PICP) levels in bronchoalveolar lavage fluid (BALF) were tested, n = 6, 3, 6, 3, respectively in each group. E-H , mice were treated intraperitoneally with integrin β3 inhibitor cilengitide 4mg/kg once a day for 3 consecutive days before being subjected to MV for 2h. Lung tissues were harvested 7days after intubation. E and F , Lac and PICP were tested in BALF, n = 6, 4, 6, 6, respectively in each group. G and H , lung tissues were stained with fluorophore-labeled antibodies against fibroblast marker FN (red), PKM2 (green) and collagen I (green), DAPI stain was used to detect nuclei (blue), original magnification × 400, scale bars correspond to 20 µm. Data are expressed as means ± SEM, *p < 0.05, **p < 0.01, *** p < 0.001 (ANOVA).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Inhibition, Staining, Marker, Labeling

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis.

doi: 10.7150/thno.72328

Figure Lengend Snippet: Figure 2. MV leads to upregulation of integrin β3 and PKM2 in pulmonary fibroblasts. Mice lung tissue were acquired at Day 7 after 2 h of MV. A, functional enrichment analysis with GO Biological Processes was performed using GOrilla with the differential genes upregulated and downregulated in pulmonary fibroblasts cluster. B, heatmaps are showing the upregulated and downregulated genes of integrin and glycolysis in pulmonary fibroblasts cluster. C, protein expression of integrin β3 and pyruvate kinase M2 (PKM2) in lung homogenates was assessed by immunohistochemistry and quantified by the average optical density value (AOD), n = 3 per group. D, protein expression of integrin β3 and PKM2 in lung homogenates was determined by western blot analysis. Relative densitometry of the protein bands of integrin β3 and PKM2 over β-actin is displayed in bar graphs, n = 3 in sham group and n = 9 in MV group. Data are expressed as means ± SEM. *p < 0.05, **p < 0.01, *** p < 0.001(t test).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Functional Assay, Expressing, Immunohistochemistry, Western Blot

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis.

doi: 10.7150/thno.72328

Figure Lengend Snippet: Figure 3. MV leads to upregulation of integrin β3 and PKM2 in pulmonary fibroblasts. Mice lung tissue were acquired at Day 7 after 2 h of MV. A and B, lung tissues were stained with fluorophore-labeled antibodies against fibroblast marker fibronectin (FN) (Alexa Fluor 568, red), integrin β3(β3) (fluorescein isothiocyanate, green) and PKM2 (fluorescein isothiocyanate, green). 4',6-diamidino-2- phenylindole (DAPI) stain was used to detect nuclei (blue). Original magnification × 400. Scale bars correspond to 20 µm. C, flow cytometry was used to detect the expression of integrin β3 and pyruvate kinase M2 (PKM2) in lung cells and in fibronectin (FN) positive fibroblasts, n = 6 per group. Data are expressed as means ± SEM. *p < 0.05, **p < 0.01, *** p < 0.001(t test).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Staining, Labeling, Marker, Flow Cytometry, Expressing

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis.

doi: 10.7150/thno.72328

Figure Lengend Snippet: Figure 7. Integrin β3 inhibition ameliorates PKM2–mediated aerobic glycolysis and MV-induced pulmonary fibrosis. A and B, both wild-type and integrin β3-deficient (β3+/-) mice were subjected to mechanical ventilation (MV) for 2 h. Lung tissues were harvested at day 7. A, lung fibrosis was quantified by Masson’s trichrome staining, original magnification × 200, scale bars correspond to 100μm, n = 6, 5, 6, 6, respectively in each group. B, integrin β3, pyruvate kinase M2 (PKM2), lactic dehydrogenase (LDHA), collagen I α1 (COL1A1) and α smooth muscle actin (α-SMA) expression was quantified by western blot analysis, n = 3 per group. C and D, mice were treated intraperitoneally with integrin β3 inhibitor cilengitide 4 mg/kg once a day for 3 consecutive days before being subjected to MV for 2 h. Lung tissues were harvested 7 days after intubation. C, lung fibrosis was quantified by Masson’s trichrome staining, original magnification × 200, scale bars correspond to 100 µm, n = 6 per group. D, integrin β3, PKM2, LDHA and COL1A1 expression was quantified by western blot analysis, n = 3 per group. Data are expressed as means ± SEM, *p < 0.05, **p < 0.01, *** p < 0.001 (ANOVA).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Inhibition, Staining, Expressing, Western Blot

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis.

doi: 10.7150/thno.72328

Figure Lengend Snippet: Figure 8. Integrin β3 inhibition ameliorates PKM2–mediated aerobic glycolysis and MV-induced pulmonary fibrosis. A-D, both wild-type and integrin β3-deficient (β3+/-) mice were subjected to mechanical ventilation (MV) for 2 h. Lung tissues were harvested at day 7. A and B, lung tissues were stained with fluorophorelabeled antibodies against fibroblast marker fibronectin (FN) (red), pyruvate kinase M2 (PKM2) (green) and collagen I (green), 4',6-diamidino-2- phenylindole (DAPI) stain was used to detect nuclei (blue), original magnification × 400, scale bars correspond to 20 µm. C and D, lactate (Lac) and type I procollagen carboxyterminal peptide (PICP) levels in bronchoalveolar lavage fluid (BALF) were tested, n = 6, 3, 6, 3, respectively in each group. E-H, mice were treated intraperitoneally with integrin β3 inhibitor cilengitide 4mg/kg once a day for 3 consecutive days before being subjected to MV for 2h. Lung tissues were harvested 7days after intubation. E and F, Lac and PICP were tested in BALF, n = 6, 4, 6, 6, respectively in each group. G and H, lung tissues were stained with fluorophore-labeled antibodies against fibroblast marker FN (red), PKM2 (green) and collagen I (green), DAPI stain was used to detect nuclei (blue), original magnification × 400, scale bars correspond to 20 µm. Data are expressed as means ± SEM, *p < 0.05, **p < 0.01, *** p < 0.001 (ANOVA).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Inhibition, Staining, Marker, Labeling