(a) Heat shock induces YAP dephosphorylation in multiple cell lines: human colon cancer HCT116; human uveal melanoma MEL270; mouse fibroblast NIH3T3; mouse squamous cell carcinoma SCC7; mouse melanoma B16-OVA. Various tumor and non-tumor cell lines were seeded onto six well plates to achieve high density (100% confluence). After 24 h, cells were heat shocked at 43°C for the indicated times and collected for YAP phos-tag gel analyses.
(b) Heat shock induced a sustained YAP dephosphorylation.
(c) Mild heat shock-induced YAP dephosphorylation is cell type-dependent. High density HEK293A and A549 cells were heat shocked at 40°C for the indicated times and collected for YAP phos-tag gel analyses.
(d) Cold stress does not affect YAP phosphorylation. High density HEK293A cells were placed at 25°C or 4°C for 30 min and 1 h; then samples were collected for YAP phos-tag gel analyses. Immunoblotting in panels a-d has been performed two times with similar results. Source data are available online.