Fig. 6
Overexpression of VEGFA by HIF-1α in GBM cells. A Staining by HE (left), RNAscope for VEGFA mRNA (middle), and IHC for HIF-1α (right) in non-Ps perinecrotic lesions of GBM. Note the dense vascular structures (indicated by arrows) in the vascular-rich area (left) and the high VEGFA mRNA and HIF-1α immunopositivity in GBM cells in the adjacent area (partitioned by dotted lines) (middle and right). Necrotic areas are indicated by asterisks and partitioned by dotted lines. Insets show the magnified views of the boxed area. Original magnification, × 100 and × 400 (inset). Scale bar = 200 μm. B Left: RT-PCR analysis of VEGFA mRNA levels in KS-1 cells after 50 and 100 μM CoCl2 treatment for 24 h. Right: KS-1 cells were transfected with VEGFA-luc reporter constructs, together with 100 and 250 ng HIF-1α. Relative activity was determined based on arbitrary luciferase light units normalized to pRL-TK activity. The activities of the reporter plus the effector relative to that of the reporter plus empty vector are shown as means ± SDs. The experiment was performed in duplicate