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. 2023 Jan 7;24(2):1221. doi: 10.3390/ijms24021221

Figure 5.

Figure 5

Effect of TNF-α, estrone and estradiol on driving stemness and EMT in HeLa pERα cells. (A) Western Blot of ERα expression using GAPDH as housekeeping. (B) Gene expression profile of ERα using qPCR, graphed as mean ± SEM (n = 3) using WT as normalizer and GAPDH as housekeeping gene (*** p < 0.001 vs Wild Type). (C) qPCR for the expression of GREB1 in WT and pERα HeLa cells treated with 10 nM E2 for 4 h (data normalized to 1 for control (EtOH) using GAPDH as the housekeeping gene). (D) ES-TF expression in HeLa pERα cells assayed using qPCR after exposure to 10 ng/mL TNF-α alone or in combination with 10 nM E1 or E2 for at least 1 week (data normalized to 1 for control using GAPDH as the housekeeping gene). (C,D) qPCR analysis for the expression of the EMT transcription factors SLUG, TWIST1 and SNAIL (E) and the EMT markers Vimentin and N-Cadherin (F) in HeLa pERα cells treated with the vehicle (EtOH, control) or 10 ng/mL TNF-α alone or in combination with 10 nM E1 or E2 for 3 weeks (data normalized to 1 for control using GAPDH as the housekeeping gene). All data are graphed as mean ± SEM from experiments performed in triplicates and repeated at least 3 times. * p < 0.05 ** p < 0.01 *** p < 0.001 vs control; # p < 0.05 ## p < 0.01 ### p < 0.001 vs TNF-α.