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. 2008 Nov 7;283(45):31183–31196. doi: 10.1074/jbc.M801606200

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Copyright © 2008, The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — GR antagonist, RU486, blocks DEX-induced up-regulation of CaD expression. A549 cells were incubated with vehicle or 1 μm DEX for the indicated times, and the expression levels of CaD mRNA (from real time qPCR) (A) and CaD protein (from Western blot) (B) are shown (mean ± S.D., ^**, p < 0.01, ^***, p < 0.001; n.s., not significant). C, A549 cells were pretreated with CHX for 2 h and incubated with vehicle or 1 μm DEX for 24 h in the presence of CHX. CaD mRNA was measured by real time qPCR. D and E, A549 cells incubated with 1 μm DEX in the presence of 10 μm RU486 or 10 μm RU28318. The expression of CaD mRNA (after 24 h (D); from real time qPCR) and CaD protein (after 48 h (E); from Western blot) are shown (mean ± S.D., ^***, p < 0.001). F, effect of RU486 or RU28318 on cell migration assayed in Transwell migration chambers. A549 cells were incubated with a combination of 1 μm DEX and 10 μm RU486 or RU28318 (mean ± S.D., ^***, p < 0.001). G, A549 cells were incubated with a combination of 1 μm DEX and 10 μm RU486 or 10 μm RU28318 for 48 h and stained with anti-CaD antibody (green in merged image) and phalloidin (red in merged image). Bar, 50 μm.