Fig. 1
From: Prion protein regulates invasiveness in glioblastoma stem cells
Characterization of U87 and U251 PrPC KO cells. (a) Illustration of the study design for the generation of PrPC KO cells. (b) RT-qPCR of PRNP mRNA amplifying the region inside the deletion site (inDEL) in U87 cells (n = 4; **P < 0.01; ****P < 0.0001). (c) Expression of PrPC protein in U87 and U251 WT and KO cells, in monolayer (M) and neurosphere (N) conditions (left) and analysis of the expression of PrPC in WT cells through band densitometry (right). Ratio between PrPC and Actin (n = 3; *P < 0.05; ***P < 0.001; ****P < 0.0001). (d) Histogram of cell surface expression of PrPC in U87 WT and KO neurosphere cells, and in U251 WT and KO monolayer cells. (e) Heatmaps depicting the relative gene expression of stem cell markers in U87 and U251 KO monolayer cells and WT and KO neurosphere cells, in relation to their monolayer WT counterparts. Asterisks (*) represent a comparison with the monolayer WT group, and plus signs (+) represent a comparison with the neurosphere WT group (p values are described in the Results section). (f) Histograms of cell surface expression of CD133 and SSEA1 proteins in U87 WT and PrPC KO neurospheres. (g) Growth curve of U87 and U251 WT and KO cells in monolayer condition (n = 6) (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 comparing WT vs. KO of the same cell line; ++++p < 0.0001 comparing U87 WT vs. U251 WT cells). (h) Self-renewal assays measuring the number of neurospheres in U87 and U251 WT and KO cells (n=*P < 0.05; **P < 0.01)