Fig. 4: p38 MAPK promotes stabilization and nuclear translocation of PDCD5 in a response to TGF-β.
From: Club cell-specific role of programmed cell death 5 in pulmonary fibrosis
a C22 cells were transfected with the indicated plasmids for 24 h and then treated with TGF-β1 for 12 h. Immunofluorescence staining was performed with a FITC-conjugated Flag antibody. Scale bars = 50 μm. b C22 cells were treated with each of the indicated inhibitors (10 μM SB203580; 10 μM TBB; and 10 μM SP600125) for 2 h before TGF-β1 (20 ng/ml) treatment. Immunoblotting with the indicated antibodies was then carried out. Representative blots from three independent experiments are shown. c, e C22 cells were treated with TGF-β1 and/or indicated inhibitors. Immunofluorescence staining was performed with anti-p-PDCD5 and anti-PDCD5 antibodies after 12 h TGF-β1 treatment. Scale bars = 50 μm. d C22 cells were treated with TGF-β1 and/or indicated inhibitors. The levels of indicated genes were analyzed by qRT-PCR. Error bars, mean ± s.e.m. (n = 3 in each group); ***p < 0.0007; ****p < 0.0001; n.s. not significant, one-way ANOVA with Tukey’s test f In vitro kinase assay was performed using GST-PDCD5 as a substrate. Error bars, mean ± s.e.m. (n = 3 in each group); ****p < 0.0001. Statistical analysis was performed with one-way ANOVA with Tukey’s post hoc test (d, f). Radioactive in vitro kinase assay was performed in GST-PDCD5 or PDCD5S119A with p38α recombinant protein. PDCD5 phosphorylation levels were analyzed by autoradiography (lower panel). Representative images and blots from three independent experiments are shown (a–c, e, f). Source data are provided in the Source Data file.
