Figure 2: PCa cell lines profiled for REST expression and characterization of EMT and stemness signature.

(A and B) LNCaP, CWR22Rv1, PC3, and DU145 were examined for REST protein by immunoblotting (A) and REST mRNA by RT-qPCR (B). n = 3. (C) Immunoblotting of protein expression for REST, NE marker SYP, and EMT and stemness markers as indicated in LNCaP and CWR22Rv1 cells. n = 3. (D) mRNA levels of NE and EMT markers and CD44 in LNCaP and CWR22Rv1 cells. n = 2. (E and F) Immunofluorescence micrographs (E) and flow cytometric analysis (F) of CD44 expression in LNCaP and CWR22Rv1 cells. n = 2. (G and H) LNCaP and CWR22Rv1 cells were seeded in transwell (1 × 104) or cultured in defined serum-free sphere medium with growth factors (2 × 104) and measured at 3 and 14 days, respectively. The number of migrated cells was quantified by the average from 5 microscopic fields (G). The number of spheres was calculated (H). Representative images (left) and quantification (right) in both PCa cell lines. n = 3. Data represent means ± SD. **p < 0.01, ***p < 0.001 by Student’s-t test.