Fig. 4
From: c-Jun N-terminal kinase activation by nitrobenzoxadiazoles leads to late-stage autophagy inhibition
Autophagic flux studies in JNK1-silenced cells. a Western blot for JNK1 in untransfected, scrambled shRNA-transfected and JNK1 shRNA-transfected cells. β-Actin was used to ensure equal loading and transfer of samples. b Western blot analysis of JNK phospho-activation in cells treated with vehicle (CTRL) or 5 μM NBDHEX for the indicated times. The filter was probed with anti-phospho-JNK (p-JNK) and anti-JNK1 antibodies; β-actin was used to ensure equal loading and transfer of samples. c Lysates from scrambled shRNA- and JNK1 shRNA-transfected cells treated with vehicle (CTRL) or 5 μM NBDHEX for 24 h, incubated or not with 100 nmol/l BAF for 3 h before lysis, were subjected to western blot for LC3-II and β-actin. d LC3-II/β-actin ratios as determined from the densitometric analysis of autoradiograms obtained in replicates of the experiment shown in c, expressed in arbitrary units (mean ± SEM, n ≥ 3; *p < 0.05 vs. CTRL; #p < 0.05 vs. minus BAF). e Autophagic flux indexes calculated from the data illustrated in d as the difference in LC3-II/actin ratios between samples plus and minus BAF (*p < 0.05 vs. CTRL), expressed in arbitrary units (a.u.)