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. 2013 Aug 26;8(8):e72053. doi: 10.1371/journal.pone.0072053

Figure 5. ZNF703-mediated activation of Akt/mTOR signaling can be inhibited by rapamycin.

Figure 5

(A) MCF-7-vector and MCF-7-ZNF703 cells were treated with DMSO, 0.1 µM rapamycin, or 1 µM rapamycin for 24 hours before cell lysis. Western blot analysis of lysates using the indicated antibodies. A GAPDH antibody was used as a loading control (B) MCF-7-vector and MCF-7-ZNF703 cells were treated with DMSO, 0.01 µM rapamycin, or 0.1 µM rapamycin for 24 hours. Western blot analysis using the indicated antibodies. (C) DMSO or various concentrations of TAM were added to three MCF-7-ZNF703 cell groups that were treated with DMSO, 0.1 µM rapamycin, or 1 µM rapamycin for 72 hours. Survival rates were examined by the SRB assay. MCF-7-ZNF703 DMSO vs. MCF-7-ZNF703 0.1 µM rapamycin, P<0.001; MCF-7-ZNF703 DMSO vs. MCF-7-ZNF703 1 µM rapamycin, P<0.01; MCF-7-ZNF703 0.1 µM rapamycin vs. MCF-7-ZNF703 1 µM rapamycin, P>0.05, paired t-test was performed. (D) Expression of p-mTOR (S2448), p-4E-BP1 (T37/46) and p-4E-BP1 (T70) in (B) were measured using ImageJ software and normalized to GAPDH levels (y-axis). *P<0.05, **P<0.01.