Fig. 4: DMRTA2 is indispensable for LN18 sphere formation and glioma stem cells driven support of angiogenesis.

A The efficacy of DMRTA2 knock-down was evaluated by qRT-PCR 48 h after siRNA transfection (n = 4 biological replicates). B Western blot showing reduction of the DMRTA2 protein level in LN18 cells after DMRTA2 knock-down. C Cell proliferation of LN18 cells decreased in DMRTA2-depleted cells as verified with BrdU incorporation test (n = 4 biological replicates in triplicates). D Viability of LN18 cells was not affected by DMRTA2 knock-down as evaluated using MTT metabolism test (n = 2 biological replicates in triplicates). E Sphere formation capacities decrease in LN18 cells depleted of DMRTA2. Representative photos of LN18-derived spheres untreated, or transfected with siCTRL or siDMRTA2 and cultured for 7 days under sphere-forming conditions, scale bar—200 µm. F Number of spheres formed by LN18 cells was reduced after DMRTA2 knock-down (n = 4 biological replicates). G Representative photos of vascular nets formed by HUVEC alone or in co-culture with control LN18 or DMRTA2 depleted cells, scale bar—200 µm. H Quantification of selected properties of the vascular net formed by HUVEC alone or in co-culture with control or DMRTA2 depleted LN18 cells (n = 4 biological replicates). UNT—control untreated cells; *p < 0.05; **p < 0.01; ***p < 0.001; Hedge’s ‘g’ stands for effect size.