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. Author manuscript; available in PMC: 2018 Jan 28.
Published in final edited form as: Nat Cell Biol. 2017 Jul 28;19(8):996–1002. doi: 10.1038/ncb3581

Figure 1. p38-mediates stress-induced TEAD cytoplasmic translocation.

Figure 1

a, Immunofluorescence staining of TEAD and YAP/TAZ in HEK293A cells treated with YAP-inhibiting signals. b, Immunofluorescence detects TEAD cytoplasmic translocation by environmental stress. c, Effect of p38 inhibitors on osmotic stress induced-TEAD cytoplasmic translocation. HEK293A cells were pretreated with p38 inhibitors, and stimulated with NaCl and stained for immunofluorescence. d, Ectopic expression of MKK3/p38 promotes TEAD4 cytoplasmic translocation. 24 hr after transfection, cells were treated with p38 inhibitors for 8hr and stained for immunofluorescence. e, Immunofluorescence staining shows deletion of p38 impairs TEAD nucleocytoplasmic shuttling by osmotic stress. Data for two independent p38 4KO clones are shown. f, Western blotting of p38 isoforms in p38 4KO cells. g, p38 mediates inhibition of YAP-TEAD target gene expression by stress. WT and TEAD KO cells were pretreated with NaCl and SB203580 as indicated, and then stimulated with 10% serum. CTGF mRNA expression was measured by qRT-PCR. Data are presented as mean ± s.e.m. from n=3 independent experiments. h, Osmotic stress inhibits YAP-TEAD target gene expression. Cells were subject to serum starvation or NaCl, and then LPA-induced CTGF and CYR61 mRNA expression was measured by qRT-PCR. Data are presented as mean ± s.e.m. from n=3 independent experiments. i, Correlation between stress-induced cytoplasmic translocation of TEAD and p38. Cells were stimulated with NaCl for the indicated times and then subjected to immunofluorescence to detect p38. Quantification of TEAD nuclear localization (N) and cytoplasmic localization (C) is provided. Random views (~100 cells) were selected for quantification. j, Time course of p38 activation by NaCl. Western blotting of phospho-p38 and its substrate phospho-MK2 upon NaCl treatment. k, Inverse correlation between stress-induced cytoplasmic translocation of TEAD and phospho-p38. Cells were stimulated with NaCl for 1 hr and then subjected to immunofluorescence using a phospho-p38 antibody. Scale bars in a–e, i, and k are 20μm. Statistics source data are shown in Supplementary Table 1. Unprocessed scans of blots are shown in Supplementary Figure 5.