Fig. 3: Effects of E2 on intranuclear AQP2 expression in tumor cell lines. | Cell Death Discovery

Fig. 3: Effects of E2 on intranuclear AQP2 expression in tumor cell lines.

From: Estrogen nuclear receptors affect cell migration by altering sublocalization of AQP2 in glioma cell lines

Fig. 3

a Experiments were repeated with two antibodies of AQP2 (DF7569 and Ab62628) in U87 cells. AQP2 was stained in red. Cell nuclei were stained with DAPI. b The phenomena of AQP2 sublocalization in the nuclei in cell lines were repeated and were confirmed in six other cell lines. c, d Western blot analysis of the cytoplasm and nucleus separately showed that E2 decreased total AQP2 expression in both the nucleus and cell membranes without change in the cytoplasm of U87 cells. c, d showed the results of RT-qPCR and western blotting, respectively. e, f E2 inhibited the total expression of p-AQP2 (ser256) and p-AQP2 (ser264) tested by western blotting and RT-qPCR, respectively. High expression level of both p-AQP2 (ser256) and p-AQP2 (ser264) observed in the AQP2 overexpression was attenuated by E2. The total expression level of p-AQP2 (ser261) was promoted by E2 and attenuated by overexpression of AQP2. ***P < 0.001 compared to control. g, h There was no obvious phosphorylation observed in the nucleus tested by western blotting and RT-qPCR, respectively. Three independent experiments were repeated. **P < 0.01 compared to control, *P < 0.05 vs. control. AQP2-membrane cell membrane, OE overexpression

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