Figure 5

siREP1-mediated cell death was alleviated with inhibition of FOXO3 in colon cancer cells. (a–e) HCT116 cells were transfected with siRNAs targeting REP1 and FOXO3 as indicated. (a) Expressions of FOXO3, REP1, BIM, BAK, and p27 were analyzed by immunoblotting. β-ACTIN was included as an internal loading control. Numbers below blots indicate the expression as measured by fold change. (b and c) Transfected cells were cultured in serum-starvation condition (0.1% FBS). (b) Number of live cells was counted by trypan blue staining. (c) The frequency of apoptotic cells (active caspase-3+) was determined by flow cytometry analysis. (d–e) Transfected cells were treated with 20 uM of 5-FU for 24 h. (d) Number of live cells was counted by trypan blue staining. (e) The frequency of apoptotic (active caspase-3+) cells was determined by flow cytometry analysis. All graphs represent two independent experiments performed in triplicate. Error bars represent S.D. from the mean. *P<0.01, **P<0.001