Figure 3.

Degradation of 14-3-3η and PDK1 activation are observed during inflammation. (a) pPDK1 and PDK1 were analyzed by western blotting in mucosal samples of C57BL/6J mice exposed to DSS (3%) for 4 days. Actin was used as a loading control. (b) pPDK1 and PDK1 protein levels were analyzed in SW480 cells transfected with 14-3-3θ, 14-3-3η, 14-3-3β, 14-3-3ɛ or 14-3-3ζ. Densitometric values for pPDK1 compared with control are displayed. Overexpressed proteins were detected with anti Oct-A (Flag) probe. Actin was used as a loading control. (c) 14-3-3η, 14-3-3θ, 14-3-3β, 14-3-3ɛ and 14-3-3ζ protein levels were analyzed in the mucosal samples of control and DSS-treated mice (C57BL/6J) by western blotting. Actin was used as a loading control. (d) Graph shows relative densitometric values of pPDK1, 14-3-3η and 14-3-3ζ obtained by western blots from mucosal cell lysates of C57BL/6J mice that were treated with DSS. Densitometric values were normalized with respect to controls (n=5). *P<0.05; **P<0.001. (e) pPDK1 (Green) and 14-3-3η (Green) were analyzed in the mucosa of control and colitic mice by immunofluorescence. C57BL/6J received drinking water or drinking water with DSS (3%) during 4 days. Nuclei, blue. Bar=20 μm. Crypt plane is marked by a discontinuous line