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Fig. 3 | Cell Division

Fig. 3

From: EZH2-inhibitor DZNep enhances apoptosis of renal tubular epithelial cells in presence and absence of cisplatin

Fig. 3

EZH2 epigenetically regulated the expression of Deptor to affect mTORC1 and mTORC2 activity. NRK-52E cells were treated with 20 μM DZNep for various times. The mRNA and protein levels of Deptor were measured by RT-qPCR (a) and western blot (b), then, we treat NAK-52E cells with different concentrations of DZNep for 24 h, the protein levels of Deptor were measured by western blot (c). The promoter of Deptor was ChIP-ed with an anti-EZH2 antibody or IgG control (d). The promoter of Deptor was ChIP-ed with an anti-EZH2 or anti-H3K27me3 antibody or IgG control in cells with or without DZNep stimulation. In the second bar, the p-value of the IgG decrease was greater than 0.05, which was not statistically significant (e). NRK-52E cells were treated with various concentrations of DZNep for 24 h. The protein level of H3K27me3 was measured, and H3 was used to verify equivalent loading (f). NRK-52E cells were transfected with three different siDeptor or negative control siRNAs for 24 h, and the protein level of Deptor was measured (g). NRK-52E cells were transfected with Deptor siRNA for 24 h, followed with 20 μM DZNep stimulation. After 24 h, the protein levels of Deptor, p ~ S6k and p ~ Akt were measured (h). Data are given as the mean ± SD of three independent experiments. *p < 0.05 versus vehicle-treated cells. # p < 0.05 versus cells treated with only DZNep

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