Fig. 6

U2AF2 bound to cARF1 and promoted glioma angiogenesis by upregulating cARF1 expression in GSCs. a, b The mRNA expression of cARF1 (a) and ARF1 (b) after U2AF2 knockdown or overexpression were detected by qPCR. c The RNA pull-down assays showed the U2AF2 protein immunoprecipitation with cARF1 as detected by western blotting. d The RIP assay was performed after U2AF2 knockdown (left) or overexpression (right), followed by qPCR to detect the enrichment of cARF1. e Relative expression levels of cARF1 in the U2AF2 knockdown GSCs treated with actinomycin D at different time points were detected using qRT-PCR. f MTS assays showed that U2AF2 overexpression or knockdown of GCM affected hBMEC viability and was reversed by cARF1 knockdown or overexpression, respectively. g The EDU assay showed that U2AF2 overexpression or knockdown of GCM affected the proliferation of hBMECs and was reversed by cARF1 knockdown or overexpression, respectively. Scale bar = 50 μm. h A representative Transwell assay showed that U2AF2 overexpression or knockdown GCM affected the invasion of hBMECs and was reversed by cARF1 knockdown or overexpression, respectively. Scale bar = 100 μm. i Representative tube formation assay showed that U2AF2 overexpression or knockdown GCM affected the tubulogenesis of hBMECs and was reversed by cARF1 knockdown or overexpression, respectively. Scale bar = 100 μm. j, k The qPCR (i) and ELISA assay (j) indicated that U2AF2 overexpression or knockdown of GCM regulated the mRNA expression and secretion of VEGFA in GSCs and was reversed by cARF1 knockdown or overexpression, respectively. EV: empty vector, OE: overexpression, NC: negative control, KD: knockdown. All data are expressed as the mean ± SD (three independent experiments). ^*p < 0.05; ^**p < 0.01; ^***p < 0.001