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. 2015 Sep 7;4(9):e167. doi: 10.1038/oncsis.2015.26

Figure 8.

Figure 8

FOXO3a is required for the anti-proliferative and epirubicin-induced cytotoxic function of FOXK2. MCF-7 cells were transiently transfected with either the control pCMV or FOXK2 and non-silencing control (NSC) siRNA or the FOXO3a siRNA smart pool. (a) Twenty-four hours after transfection, protein lysates were prepared from these cells and then analysed for the expression of FOXK2, FOXO3a and β-tubulin (b) The transfected cells were also treated with a range of doses of epirubicin (0–5000 nm) and their proliferative rates assayed by SRB assay at 48 and 72 h after epirubicin treatment. Statistical significance was determined by Student's t-test (two-sided; *P⩽0.05, **P⩽0.01, NS, not significant) by comparing the proliferation rates of cells transfected with pFOXK2 with the control pCMV transfected cells. (c) Twenty-four hours after transfection, the 2000 cells were seeded in six-well plates, treated with 0, 10 or 20 nm of epirubicn, grown for 15 days and then stained with crystal violet. Two representative independent experiments are shown (top panel). (d) The result (bottom panel) represents the average of three independent experiments±s.d. Statistical significance was determined by Student's t-test (two-sided; *P⩽0.05, **P⩽0.01, NS, not significant).