Figure 4
From: Tumor suppressor FOXO3 participates in the regulation of intestinal inflammation
TNFα-induced inactivation of FOXO3 is controlled by IKK. (a) Total proteins from untreated and TNFα-treated cells were separated on SDS-PAGE and immunoprobed with an antibody against phosphorylated FOXO3 at the Ser644 IKK-dependent position. Immunoblots were also re-probed with an antibody against actin. (b) The HT-29 monolayers were pre-treated with the IKK inhibitor, PS1145, and induced with TNFα for various time points. Protein was separated on SDS-PAGE and immunoprobed with an antibody against total FOXO3 and actin. The graphs represent the densitometric analysis showing a significant decrease of FOXO3 (*) after TNFα treatment (n=3, P<0.05) and protection of degradation with the IKK inhibitor. (c) Protein from the monolayers pretreated with PS1145 and TNFα was separated and immunoprobed against phosphorylated FOXO3 at Thr32 PI3K-dependent site and actin. The densitometric analysis shows a significant increase (*) in phosphorylated FOXO3 after TNFα treatment, which was not attenuated in the presence of PS1145.
