cilengitide treatment  (MedChemExpress)

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Thy-1 depletion and integrin β3 upregulation-mediated PI3K-Akt-mTOR pathway activation inhibits lung fibroblast autophagy in lipopolysaccharide-induced pulmonary fibrosis

doi: 10.1038/s41374-019-0281-2

Figure Lengend Snippet: Integrin β3 inhibition precludes LPS-induced PI3K-Akt-mTOR activation and lung fibroblast autophagy blockade. Western blot was used to detect the expression of integrin β3 ( a ), phospho-AKT (p-AKT), total AKT, phospho-mTOR (p-mTOR) and total mTOR ( b ). Representative images showing protein expression of LC3 I, LC3 II and P62 in MRC-5 cells challenged without or with 1 μg/ml LPS in the absence or presence of the inhibitor cilengitide (2 mM) for 24 h ( c ). Quantitation of p-AKT/total AKT, p-mTOR/total mTOR, LC3 II/ I, integrin β3 and P62 protein levels normalized to GAPDH ( d ). Lung fibroblasts were assessed by transmission electron microscopy. White arrows indicate autophagosomes ( e ), the semi-quantification of autophagosomes of per cell were also shown ( f ). Values are mean ± SD from triplicate experiments. * p < 0.05 vs control group; ** p < 0.01 vs control group; # p < 0.05 vs LPS group; ## p < 0.05 vs. LPS group

Article Snippet: MRC-5 cells in the logarithmic growth phase were seeded into 6-well plates at a density of 2 × 10 5 cells/mL (2 mL in each well), and stimulated with 1 μg/ml LPS to generate an autophagy inhibition model. Then, treatment with cilengitide (#S7077, Selleck, an integrin β3 inhibitor), integrin β3 knockdown lentivirus (Itgb3-KD), Thy-1 knockdown lentivirus (Thy-1-KD) and Thy-1 overexpression lentivirus (Thy-1-OE) (Genomeditech, Shanghai, China) were used to inhibit or overexpress integrin β3 or Thy-1 at the protein and gene levels.

Techniques: Inhibition, Activation Assay, Western Blot, Expressing, Quantitation Assay, Transmission Assay, Electron Microscopy, Control

Journal: Laboratory Investigation; a Journal of Technical Methods and Pathology

Article Title: Thy-1 depletion and integrin β3 upregulation-mediated PI3K-Akt-mTOR pathway activation inhibits lung fibroblast autophagy in lipopolysaccharide-induced pulmonary fibrosis

doi: 10.1038/s41374-019-0281-2

Figure Lengend Snippet: Integrin β3 inhibition prevents LPS-induced autophagy inhibition and LPS-induced pulmonary fibrosis. Male C57BL/6J mice aged 6–8 weeks ( n = 6/group) were pretreated with cilengitide (2 mg/kg), followed by LPS (5 mg/kg) administration for consecutive 5 days. Western blot was performed to detect the expression levels of integrin β3, LC3, P62 ( a ) and α-SMA ( b ) in the lung tissue. The severity of collagen deposition was measured by assessing hydroxyproline and collagen amounts ( c ). The severity of pulmonary fibrosis was determined by hematoxylin-eosin (H&E) staining; collagen deposition was revealed by Masson’s trichrome staining, and α-SMA expression in the lung tissue was detected by immunohistochemistry (magnification, ×200) ( d ). Values are mean ± SD ( n = 6). * p < 0.05 vs. control group; ** p < 0.01 vs. control group; # p < 0.05 vs. LPS group; ## p < 0.05 vs. LPS group

Article Snippet: MRC-5 cells in the logarithmic growth phase were seeded into 6-well plates at a density of 2 × 10 5 cells/mL (2 mL in each well), and stimulated with 1 μg/ml LPS to generate an autophagy inhibition model. Then, treatment with cilengitide (#S7077, Selleck, an integrin β3 inhibitor), integrin β3 knockdown lentivirus (Itgb3-KD), Thy-1 knockdown lentivirus (Thy-1-KD) and Thy-1 overexpression lentivirus (Thy-1-OE) (Genomeditech, Shanghai, China) were used to inhibit or overexpress integrin β3 or Thy-1 at the protein and gene levels.

Techniques: Inhibition, Western Blot, Expressing, Staining, Immunohistochemistry, Control

Journal: Cancer research

Article Title: Antagonizing integrin beta3 increases immune suppression in cancer

doi: 10.1158/0008-5472.CAN-15-2663

Figure Lengend Snippet: (A) Experimental schema. 1×105 PyMT-BO1-GFP-Luc cells were injected into MFP of 8-week-old female WT mice (n=5). Starting at day 10 after tumor cell injection, mice were treated daily with cilengitide for 5 days. At day 14, all mice were sacrificed. (B) Tumor growth was measured at the indicated time points by BLI. (C) Tumor weight at day 14. (D) TAM populations in tumor tissue were analyzed by FACS. Data represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: For the cilengitide treatment, mice were given cilengitide (5mg/kg, Selleckchem) by intraperitoneal injection for the indicated time point.

Techniques: Injection

Journal: Cancer research

Article Title: Antagonizing integrin beta3 increases immune suppression in cancer

doi: 10.1158/0008-5472.CAN-15-2663

Figure Lengend Snippet: (A) Western blots of p-SYK and SYK in LPS-stimulated WT and Itgb3−/− BMMs. (B) Western blots of p-STAT1 and STAT1 in LPS-stimulated WT and Itgb3−/− BMMs. (C) Western blots of p-STAT1 and STAT1 in LPS-stimulated WT BMMs with or without cilengitide pretreatment. (D) Western blots of p-STAT1 and total STAT1 in WT BMMs overexpressing integrin beta3. M+pMX: macrophage treated with empty pMX vector. M+hβ3: macrophage treated with human integrin beta3 constructed pMX vector. (E) Western blots of p-STAT6 and total STAT6 in IL-4 stimulated WT and Itgb3−/− BMMs. (F) Ym1 mRNA expression after IL-4 treatment of WT and Itgb3−/− BMMs. (G) Itgb3 mRNA expression level after IL-4 treatment in Stat6−/− macrophages. (H) Western blot analysis of integrin beta3 expression in WT, Itgb3−/−, and Stat6−/− BMMs after 5ng/ml IL-4 treatment for 48 hours.

Article Snippet: For the cilengitide treatment, mice were given cilengitide (5mg/kg, Selleckchem) by intraperitoneal injection for the indicated time point.

Techniques: Western Blot, Plasmid Preparation, Construct, Expressing

Journal: Cancer research

Article Title: Antagonizing integrin beta3 increases immune suppression in cancer

doi: 10.1158/0008-5472.CAN-15-2663

Figure Lengend Snippet: (A) PyMT-BO1-GFP-Luc MFP tumor established on β3KOM mice (n=5). From day 10 after tumor cell injection, mice were treated daily with cilengitide for 5 days. Tumor growth was measured at the indicated time points. (B) Experimental schema. 1×105 PyMT-BO1-GFP-Luc cells were injected into MFP of 8-week-old female WT mice at day 0. Anti-CSF1 antibody treatment started on day 6 by i.p. injection of 1mg of antibody per mouse, followed by 0.5mg of antibody on day 9 and day 12. Cilengitide treatment started on day 9 for 5 days with a dosage of 5mg/kg per mouse by i.p. injection (n=7 or 8 per group). (C) Tumor burden was measured by BLI at day 15. (D-G) Tumor-infiltrating myeloid cells and T-cells were measured by FACS at day 15. Data represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: For the cilengitide treatment, mice were given cilengitide (5mg/kg, Selleckchem) by intraperitoneal injection for the indicated time point.

Techniques: Injection