Fig. 6

PDZ-binding motifs are required for TPM2 function. A Co-IP was performed using cell lysates obtained from HEK293T cells co-transfected with Myc-PDLIM7 and Flag-TPM2 or Flag-TPM2ΔC. B Left panel: staining of recombinant GST-TPM2 and GST-TPM2ΔC fusion proteins by Coomassie brilliant blue. Right panel: the GST-pulldown assays were carried out by incubating GST-TPM2 or GST-TPM2ΔC bound beads with Myc-PDLIM7-expressing HEK293T cell lysates and analyzed by Western blot. C–G TPM2ΔC was overexpressed to explore its function in C4-2B and PC-3 cells. The proliferation capacities of the PCa cells were detected by CCK-8, colony formation, and EdU assays. Transwell assays were used to assess the migration and invasion ability of cells. C–E Cell proliferation assays: CCK-8 and EdU assays. F, G Transwell migration and invasion assays. H The level of YAP1 in the nucleus and cytoplasm after TPM2ΔC overexpression. I Immunofluorescence assay was used to detect the distribution and expression level of YAP1 in C4-2B and PC-3 cells after overexpressing TPM2ΔC. J The expression levels of YAP1 and p-YAP1(S127) in C4-2B and PC-3 cells after overexpression of TPM2ΔC. K The expression level of YAP1 target genes: c-MYC, AREG, and PCNA. L, M Effect of TPM2 on the protein stabilization of YAP-1 in cells treated with 50 µM cycloheximide (CHX) for the indicated times. N Tumor growth curves of PC-3 cells express indicated protein in a xenograft mouse model. O Images of excised tumors. P Tumor weights. ns, no significance