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. 2021 Jun 30;9:675998. doi: 10.3389/fcell.2021.675998

FIGURE 4.

FIGURE 4

6C can suppress the occurrence of EMT. (A) Immunofluorescence staining of EMT-related genes after 1 week of cell culture. ZEB1 and Snail are transcription factors of EMT, β-catenin and α-SMA are markers of EMT (green). Nuclei were stained with DAPI (blue). The expression of EMT-related genes in group 6C was significantly reduced. (B) qRT-PCR analysis of ZEB1/2, Snail, β-catenin and α-SMA gene expression (normalized to ctrl) after 1 week of cell culture. (C) Comparison of cell morphology. a, Treat with 6C for 1 week. b, Treat with 6C for 3 days, then remove 6C and continue to cultivate for 4 days. (D) Comparison of cell morphology. a’, It was first treated with 6C for 3 days, and then cultured with 6C plus 10% FBS for another 4 days. b’, Treat with 6C for 3 days, then remove 6C and continue to incubate with 10% FBS for 4 days. (E) Statistical analysis of cell diameter after C and D treatment. (F) After C treatment, qRT-PCR analysis of P63, K14, K12, Pax6, Ki67, and α-SMA gene expression (normalized to 6C). (G) After D treatment, qRT-PCR analysis of P63, K14, K12, Pax6, Ki67, and α-SMA gene expression (normalized to 6C plus 10% FBS). Data expressed as the means ± SEM from three separate experiments (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns: no significance).