Fig. 5
Cellular prion protein (PrP C) promotes GSC self-renewal binding HOP and modulates cell surface adhesion molecule stability. Neurosphere number (a) or size (b) after 1 week treatment every 48 h with Hsp70/90 organizing protein (HOP) and peptides pepHOP230–245 and pepHOP422–437 (1 μM), combined or alone (500nM), compared to control (n = 6, *p < 0.05, ANOVA followed by Tukey post-hoc test). c Dot plot of E-cadherin expression in parental and shRNA-PrP2 neurospheres. E-cad+ cells shown in red and E-cad– cells shown in black. d Dot plot of E-cadherin and PrPC expression in parental and shRNA-PrP2 neurospheres. e Immunofluorescence for E-cadherin (green) in parental and shRNA-PrP2 neurospheres, showing expression on the cell surface (parental) and in the perinuclear region (shRNA-PrP2). Nuclei (TO-Pro) stain shown in red. f PrPC (green) and β-catenin (red) expression and co-localization (yellow) of parental and shRNA-PrP2 neurospheres. g Migration assay, ratio between cell migration distance (halo), and neurosphere size for parental and shRNA-PrP2 neurospheres 24 h after plating on laminin-1 (n = 3, *p < 0.05). h Cell scratch assay; images of three experimental replicates were acquired and the distance of each scratch closure after 24 h was measured by comparing with the images at time 0 h for parental and shRNA-PrP2 neurospheres plated on laminin-1 (n = 4, *p < 0.05). i Dot plot of α6 integrin and PrPC expression in parental and shRNA-PrP2 neurospheres. j Immunofluorescence for β1 integrin (green) of parental and shRNA-PrP2 neurospheres. Nuclei (TO-PRO) stain shown in red. k. PrPC (green) and β1 integrin (red) expression and co-localization (yellow) of parental and shRNA-PrP2 neurospheres. Nuclei (TO-PRO) stain shown in blue; a higher magnification is shown in the inset