Figure 8: TNF signalling in Pellino3−/−-deficient MEFs.
From: Pellino3 targets RIP1 and regulates the pro-apoptotic effects of TNF-α
MEFs were isolated from Pellino3+/+ and Pellino3−/− embryos and treated in the absence (control) or presence of murine TNF (20 ng ml−1) for the indicated time periods. (a) Cell lysates were prepared and immunoblotted for expression levels of phosphorylated IκBα (p-IκBα), JNK (p-JNK), P38 (p-P38), ERK (p-ERK) and total levels of IκBα, JNK, P38 and ERK. Lysates were also probed for levels of β-actin to act as loading controls. (b) Conditioned media from treated cells were assayed for levels of IL-6 by ELISA. ***, P<0.001 (two-way ANOVA). (c) Cell lysates were probed by immunoblotting for expression levels of Bcl-x and β-actin. (d,e) mRNA expression levels of (d) A20 and (e) c-FLIP were measured by quantitative RT-PCR and expressed relative to expression levels in unstimulated Pellino3+/+ MEFs. Data represent the mean +/− s.e.m. of three independent experiments; **P<0.01 (two-way ANOVA). Full-length images of immunoblots are shown in Supplementary Fig. S15.
