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. 2017 Apr 10;7(7):1890–1900. doi: 10.7150/thno.19135

Figure 3.

Figure 3

TAM activated Nrf2 specifically in endometrial cells. A, Nrf2 expression in RL95-2, AN3CA or MCF7 cells incubated with or without 4-OH-TAM (15 μM) for 24 hours were analyzed by western blotting. B, RL95-2, AN3CA or MCF7 cells were incubated with or without 4-OH-TAM (15 μM) for 24 hours. Nrf2 distributions in nuclear and cytoplasmic extracts were explored by western blotting. GLS1 was used as the cytoplasmic protein marker while H3 (Histone H3) was used as the nuclear protein marker. C, NQO1 mRNA level in RL95-2, AN3CA or MCF7 cells incubated with or without 4-OH-TAM (15 μM) for 24 hours were analyzed with qRT-PCR. The experiments were repeated three times. Representative results from one triplicated experiment were shown as Mean±SD. Student's T test was used for the statistical analysis. The asterisk indicates the statistical significance and the octothorpe stands for no significant difference. D, Forty-eight hours after transfection of SQSTM1 siRNAs, 4-OH-TAM (15 μM) was added for another 24 hours, the effect of 4-OH-TAM on Nrf2 expression in RL95-2 and AN3CA cells with or without SQSTM1 knockdown were explored by western blotting. E, Forty-eight hours after transfection of Flag-SQSTM1/Vector, Nrf2 expression in RL95-2 (left) or AN3CA (right) cells were detected by western blotting. F, Nrf2 distribution in nuclear and cytoplasmic extracts of RL95-2 (left) or AN3CA (right) cells with or without Flag-SQSTM1 over-expression were detected by western blotting.