Fig. 2: PrPC is necessary for EMT in LUAD cell lines.
A qRT-PCR analysis of the expression of the EMT TF SNAI1, SNAI2, TWIST, ZEB1, ZEB2 in PRNP-silenced versus control H1975 cells. Western blot analysis of the expression of ZEB1 (B) and SLUG (C) in PRNP-silenced versus control H1975 cells. D GSEA analysis highlighting the EMT signature as one of the most affected pathway in H1975 cells in response to PRNP silencing. E Immunofluorescence images showing PrPC and F-actin staining in PRNP-silenced versus control H1975 cells. Analysis with the CASY cell counter revealing a decrease in proliferation (F) and an increase in cell volume (G) in PRNP-silenced versus control H1975 cells. Cell index measurements of PRNP-silenced versus control H1975 cells in xCELLigence migration (H) or invasion (I) assay. qRT-PCR analysis of the expression of the EMT TF SNAI1, SNAI2, ZEB1, ZEB2 (J) and EMT genes VIM, CDH1 and CDH2 (K) in HCC827 cells exposed to siRNA against PRNP and recombinant TGFβ1. L Western blot analysis of the expression of SNAIL, ZEB1, ZEB2, Vimentin, E-cadherin and N-cadherin in HCC827 cells exposed to siRNA against PRNP and recombinant TGFβ1. Results are expressed as means ± s.e.m of n = 2 independent triplicates (A–C, J) or n = 1 triplicate (F–I, L) of cell preparations. *p < 0.05, **p < 0.01 ***p < 0.001 vs. control (siScramble), #p < 0.05, ###p < 0.001 vs. TGFβ1 treated, siScramble cells, §p < 0.05 vs. TGFβ1 untreated, PRNP-silenced cells. Protein levels in western blots were normalized to α-tubulin (α-tub) with quantifications summarized in Supplementary Fig. S2F.
