Recombinant PEDF promotes stemness and self-renewal of GSCs.

<p>(A) Sphere formation assay was performed in X02 cells treated with recombinant PEDF (rPEDF) (100 ng/ml) or control vehicle. Images are representative of three independent experiments. The graph represents average proportion of sphere number. Counted sphere size is greater than 100 μm. All error bars represent mean ± SEM (<i>n</i> = 3). * <i>p</i> < 0.05. (B) IB analysis of Sox2, Nestin, and GFAP in X02 cells treated or not treated with rPEDF (100 ng/ml). β-actin was used as a loading control. (C) Sphere formation assay was performed in X01 cells cultured in serum-free GSC medium (containing EGF and bFGF) or serum-free GSC medium without EGF and bFGF. X01 cells cultured in serum-free GSC medium without EGF and bFGF were treated with rPEDF (100 ng/ml) or control vehicle. Images are representative of three independent experiments. The graph represents the average proportion of sphere number. Counted sphere size is greater than 100 μm. All error bars represent mean ± SEM (<i>n</i> = 3). ** <i>p</i> < 0.01. (D) IB analysis of PEDF (in medium), Sox2, Nestin, and GFAP in X01 cells cultured in three different conditions (C). α-tubulin was used as a loading control. All images were taken at 5x magnification.