Figure 4. miR-210 in Hypoxic^EXO increase EWS sphere formation.

(A) Sphere assay of A673 and SK-ES-1 cells grown under normoxic and hypoxic conditions. Quantification of spheres was performed (magnification, ×4) (mean + SEM, n = 3, ^* P ≤ 0.02, ^** P < 0.008). (B) Analysis of miR-210 expression levels in normoxic and hypoxic A673 and SK-ES-1 spheres using qRT-PCR (Mean + SEM, n = 3, ^* P ≤ 0.5). RNU6B was used as a control to normalize miRNA expression. (C) Sphere assay quantifying A673 and SK-ES-1 cells transfected with either miR-scramble or miR-210 inhibitors under normoxic and hypoxic conditions. (Mean + SEM, n = 3, ^* P ≤ 0.0001, ^** P ≤ 0.005). (D) Sphere assay quantifying A673 and SK-ES-1 cells transfected with miR-scramble, miR-210 mimics, miR-210 inhibitors or 20 μg/ml hypoxic exosomes. (Mean + SEM, n = 3, ^* P ≤ 0.002, ^** P ≤ 0.0001).