Fig. 1
From: Identification of a novel anoikis signalling pathway using the fungal virulence factor gliotoxin
MKK4 and MKK7 are required for GT-induced anoikis. a Western blot analysis of total extracts of human bronchial epithelial cells (BEAS-2B) showing increased phosphorylation of MKK4 (Ser257/Thr261) (pMKK4) and MKK7 (Ser271/Thr275) (pMKK7) as well as PARP cleavage (PARP/cPARP) after GT treatment for 4 and 6 h. b Western blot analysis showing increased phosphorylation of JNK (T183/Y185) (pJNK) and Bim (T112/S114) (pBim) and enhanced processing of caspase-3 and PARP in total extracts of WT MEFs treated with GT for 4 and 6 h. None of these changes were seen in the extracts of non-treated (NT) cells or MEFs deficient for both Mkk4 and Mkk7 (Mkk4−/−/Mkk7−/−). c, d MEFs deficient for either Mkk4 (Mkk4−/−), Mkk7 (Mkk7−/−) or both Mkk4/Mkk7 (Mkk4−/−/Mkk7−/−) exerted reduced caspase-3/7 activity (c) and annexin V-FITC staining (d) and as compared to wild-type (WT) cells when treated with GT for 6 h. e Schematic representation of how GT activates MKK4/MKK7 (MKK) and triggers JNK/Bim-mediated anoikis. Tubulin was used as loading control in a and b. Graphs in c and d show the means of at least three independent experiments ± s.e.m.; p-values: *0.05–0.01, **0.01–0.001, ***<0.001; two-way ANOVA, post hoc: Bonferroni compared to WT
