Fig. 2: CD5L is upregulated through hypoxia-induced PPARG overexpression.
From: Overcoming adaptive resistance to anti-VEGF therapy by targeting CD5L
A, B CD5L mRNA (A) and protein expression (B) in RF24 endothelial cells containing PPARG overexpressing plasmid versus empty vector (EV). C CD5L promoter constructs activation using RF24 endothelial cells containing PPARG overexpressing plasmid versus EV. D, E PPARG and CD5L mRNA (D) and protein (E) expression in RF24 endothelial cells treated with siPPARG versus siControl. F CD5L promoter construct activation using RF24 endothelial cells treated with siPPARG versus siControl. G Luciferase expression in RF24 endothelial cells after co-transfection of PPARG overexpressing plasmid and CD5L promoter construct harboring mutated PPARG binding site. H, I PPARG and CD5L mRNA (H) and protein (I) expression in RF24 endothelial cells cultured in hypoxic or normoxic conditions. J, K PPARG and CD5L mRNA (J) and protein (K) expression in RF24 endothelial cells treated for 6 and 30 h with cobalt chloride (HIF1α stabilizer). Western blots were performed from two independent technical replicates; in each repeat, the blotting, including loading control, was performed using the same sample processing controls (B, E, I, K); L PPARG and CD5L mRNA expression in RF24 endothelial cells treated with YC-1 or topotecan under hypoxic conditions. M CD5L WT promoter construct activation in RF24 endothelial cells cultured in hypoxic and normoxic conditions. N Chromatin immunoprecipitation (ChIP) analysis of the CD5L promoter using an anti-PPARG antibody under hypoxic and normoxic conditions. Data represented as mean values ± SD, determined by two-tailed Student’s t-test for A, C, F, G, H, M; one-way ANOVA Tukey’s multiple comparisons for J and L; two-way Anova Tukey’s multiple comparisons for D and N; n = 3 for D, G, H, J left panel, M and N; n = 4 for A, C, and L left panel; n = 5 for F; n = 2 for J right panel and L right panel.
