Figure 3

TERT-siSFRP1 cells exhibit a gene expression profile that is consistent with partial EMT. A, TERT-pSUPER and TERT-siSFRP1 were immunostained with either anti-E-cadherin or anti- vimentin and counterstained with DAPI. B, For each cell line, total RNA was isolated from three separate harvests for real-time PCR analysis of E-Cadherin, vimentin, ZEB1 and ZEB2. C, The SFRP1-pCDNA3.1 construct was transfected into TERT-siSFRP1 cells and RNA was isolated from 3 separate harvests. Real-time PCR analysis was performed to measure the relative expression of SFRP1, E-cadherin, vimentin, ZEB1 and ZEB2. All Real-time PCR results are from two separate experiments performed in triplicate and results were normalized to amplification of GAPDH mRNA. Bars represent mean ± SEM and are expressed as fold change with respect to TERT-pSUPER cells. *p < 0.05 ***p < 0.001 (significantly different from TERT-pSUPER cell line using student's t-test), ^‡p < 0.001 (significantly different from TERT-siSFRP1 cell line using Bonferoni's t-test following one-way ANOVA). Immunostain images were captured at 40× magnification (scale bar 50 μm).