Figure 3

CAV1 and CAVIN1 Are Direct YAP/TAZ-TEAD Target Genes

(A) Western blots of lysates from shTEADs and shCon HEK293A cells.

(B) Mixed cell population of shTEADs and shCon HEK293A cells labeled for DAPI (blue), CAV1 (red), and TEAD1 (green). Discrete cell populations are shown in Figure S4A (see also Figure S4B).

(C) Mixed cell population of shTEADs and shCon HEK293A cells labeled for DAPI (blue), TEAD1 (red), and CAVIN1 (green). Discrete cell populations are shown in Figure S4D.

Arrows in (B) and (C): examples of shTEADs cells. Scale bars in (B) and (C) represent 15 μm.

(D and E) Dot plot of CAV1 levels from images, as shown in (B) (Figure S4C; D) and dot plot of CAVIN1 levels from images, as shown in (C) (Figure S4E; E). Each dot represents one cell. Means ± SEM.

(F) qPCR data from cells as in (A) (related to Figure S4F). Means ± SD.

(G) YAP drives CAVIN1 and CAV1 promoter activity. Luciferase reporters were generated carrying either a short (−500 to +200 bp), or long (−1,907 to +200 bp) fragment of the CAV1 promoter region or a fragment (−1,250 to +150 bp) of the CAVIN1 promoter region (related to Figure S4G). The reporters were introduced into Y/T KO HEK293A cells together with either a YAP or a vector control plasmid. Note that only the long CAV1 form contains the predicted TEAD recognition motifs. Means ± SD.

(H) Real-time PCR analysis of TEAD1 chromatin immunoprecipitation (ChIP) in HEK293A cells. The precipitated DNA was quantitated using primers specific for a promoter region or a control in-gene (Ing) region. Data are means ± SD of triplicates from a representative experiment. Endogenous TEAD1 binds to both CAV1 and CAVIN1 promoters.

(I and J) YAP ChIP for CAV1 and CAVIN1 in shCon HEK293A cells (I) and in shTEADs HEK293A cells (J). Endogenous YAP binds to both CAV1 and CAVIN1 promoters in a TEAD-dependent manner.