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. 2017 Feb 7;114(8):E1500–E1508. doi: 10.1073/pnas.1612991114

Fig. 4.

Fig. 4.

SGK3 maintains EnR homeostasis. (A) Transmission electron microscopy of LET-R cells transfected with siRNA negative control or SGK3 siRNA for 72 h. (Magnification: 1,100×.) (Scale bar: 4 μm.) (B) Immunofluorescence microscopy of AI-resistant cells transfected with SGK3 siRNA or siRNA negative control. EXER-R and LET-R cells were transfected with siRNA negative control or SGK3 siRNA for 72 h and then fixed and immunostained with anti-calnexin (CNX) or anti-calreticulin (CRT) plus anti-LAMP2. After immunostaining, the cells were mounted in DAPI solution and imaged under a confocal microscope. DAPI stained nuclei blue. CNX and CRT were shown in red, and LAMP2 was shown in green. (Scale bar: 5 μm.) LAMP2 is a membrane protein of lysosome and late endosome, and thus serves as a marker of lysosome and later endosome. (C) Western blotting analysis of EXE-R and LET-R cells after being transfected with siRNA negative control or SGK3 siRNA for 72 h. (D) Western blotting analysis of EXE-R and LET-R cells after being treated with the increasing concentrations of GSK650394 for 48 h. (E) Western blotting analysis of MCF7aro cells cultured in regular growth medium or in hormone-depleted medium for 3 d. (F) Western blotting analysis of MCF7aro/TO/SGK3 cells grown in different conditions. MCF7aro/TO/SGK3 cells were cultured in normal growth medium or hormone-depleted medium and treated with or without 250 ng/mL DOX for 3 d. (G) Western blotting analysis of MCF7aro/TO/SGK3 cells grown in the medium with T plus letrozole in the presence or absence of 250 ng/mL DOX for 14 d.