Fig. 6: Inhibition of PDGFRA pathway attenuates effect of Endocan on GBM cells.

a Western blot analysis of 1079 GBM cells after lentiviral mediated knockdown of PDGFRA with 4 different shRNAs against PDGFRA or Non Target shRNA (NT) as a control. Representative result of n = 3 biological replicates. b Effect of rEndocan (10 ng/ml) on proliferation of 1079 cells infected with shNT or shPDGFRA lentiviruses. n = 3 biological replicates. ***P = 0.0004 for shNT (rEndocan vs. Control), P = 0.7498 for 3shPDGFRA (rEndocan vs. Control) and P = 0.2366 for 4shPDGFRA (rEndocan vs. Control), unpaired two-tailed Student’s t test. c Western blot analysis of 1079 cells depicting PDGFRΑ, MYC, and β-actin protein level expression in cells treated with rEndocan (10 ng/ml), rPDGF-BB (10 ng/ml), ponatinib (1 μM) and their combinations for 72 h, Representative result of n = 3 biological replicates. d Effect of ponatinib(1μM) on proliferation of 1079 cells incubated with rEndocan or rPDGF-BB (10 ng/ml). n = 3 biological replicates, one-way ANOVA followed by Dunnett’s multiple comparisons test. ***P = 0.0001 and ****P < 0.0001. e Kaplan–Meier survival analysis of Bl/6 mice intracranially injected with WTD and KOD cells and treated with 50 mg/kg/day dose of ponatinib for 5 days using oral gavage. n = 5 mice per group, P value was determined by log-rank test; **P = 0.00184 for WT control vs. WT ponatinib, P = 0.87 for Esm1 KO control vs. Esm1 KO. Ponatinib. f Representative microscopic images of immunohistochemical (IHC) staining with anti-CD31 antibody of tumor sections obtained from mice as in “E”. g Blood vessels density (n = 18 vessels/group) from tumor sections (n = 3 mice) stained with CD31 antibody was analyzed using ImageJ software. ****P < 0.0001 for WT Ponatinib vs. WT Control and P = 0.7852 (not significant) for KO Ponatinib vs. KO Control. ****P < 0.0001 for WT control vs. KO Control groups. one-way ANOVA followed by Dunnett’s multiple comparisons test. Scale bar, 100 µm. Source data are available in the Source Data file.