Fig. 7 | Nature Communications

Fig. 7

From: Identification of a novel anoikis signalling pathway using the fungal virulence factor gliotoxin

Fig. 7

Induction of integrins by PMA confers sensitivity to anoikis. a Phase contrast microscopy analysis of K562 cells treated with 10 ng/ml PMA for 96 h before adding 1 μM GT for 6 h. While untreated (NT) cells grow in suspension, PMA-treated cells adhere to the dish and become sensitive to GT-induced detachment. Scale bar = 100 µm. b GT induced caspase-3/-7 activity (taken as 100%) in PMA-treated adherent K562 cells (+PMA) but not in undifferentiated suspension cells (−PMA). c, d FACS analysis of K562 cells showing that PMA increased the surface expression of integrin β3 (c) and an active form of integrin β1 (using the anti-active integrin β1 12G10 Alexa Fluor® 488) (d). Iso: respective Ig control-FITC antibody. e Western blot analysis of total extracts of undifferentiated (−PMA) and differentiated (+PMA) K562 cells either untreated (NT) or treated with 1 µM GT for 6 h showing that PMA induced the expression of integrin (INTG) αV, β1 and β3. GT triggered the phosphorylation of MKK4, JNK and Bim (as evidence by a gel shift) as well as caspase-3 processing, but only in PMA-treated cells. Tubulin was used a loading control. Graphs in bd show the means of at least three independent experiments ± s.e.m.; p-value: ***<0.001, two-way ANOVA, post hoc: Bonferroni compared to non-treated (NT)

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