Fig. 5

TPM2 blocks PDLIM7-mediated nuclear translocation of YAP1. A, B Whole-cell lysates from C4-2B and PC-3 cells immunoprecipitated with PDLIM7 or YAP1 antibody followed by Western blot analysis. C The level of YAP1 in the nucleus and cytoplasm after PDLIM7 overexpression. D Immunofluorescence assay was used to detect the distribution and expression level of YAP1 in C4-2B and PC-3 cells after overexpressing PDLIM7. E The expression levels of YAP1 and p-YAP1(S127) in C4-2B and PC-3 cells after overexpression of PDLIM7. F HEK293T cells were transfected with HA-Ub and Flag-YAP1 with or without Myc-PDLIM7. After treatment with MG132 (10 µM, 6 h), ubiquitination of YAP1 was measured. G, H Effect of PDLIM7 on the protein stabilization of YAP-1 in cells treated with 50 µM cycloheximide (CHX) for the indicated times. I The expression level of YAP1 target genes: c-MYC, AREG, and PCNA. J The stain of YAP1 in PDLIM7-overexpressing subcutaneous tumors. Scale bars represent 50 μm. K Protein structure of TPM2, PDLIM7, and YAP1. L The level of YAP1 in the nucleus and cytoplasm after TPM2 overexpression. M Immunofluorescence assay was used to detect the distribution and expression level of YAP1 in C4-2B and PC-3 cells after overexpressing TPM2. N The expression levels of YAP1 and p-YAP1(S127) in C4-2B and PC-3 cells after overexpression of TPM2. O HEK293T cells were transfected with HA-Ub and Flag-YAP1 with or without Myc-TPM2. After treatment with MG132 (10 µM, 6 h), ubiquitination of YAP1 was measured. P, Q Effect of TPM2 on the protein stabilization of YAP-1 in cells treated with 50 µM cycloheximide (CHX) for the indicated times. R The expression level of YAP1 target genes: c-MYC, AREG, and PCNA. S The stain of YAP1 in TPM2-overexpressing subcutaneous tumors. Scale bars represent 50 μm