Figure 5.

YAP Mediates the Effect of CAV1 in ECM Remodeling

(A and B) Collagen gel retraction induced by MEFs transfected with YAP-Flag or YAP(S5A)-FLAG.

(A) Cells were embedded in 3D collagen gels, and images were acquired 72 hr later to monitor gel retraction. Collagen organization was determined by second harmonic generation (SHG) microscopy (black and white images). Mock represents mock transfection.

(B) Corresponding ImageJ quantification of gel contraction, measured as the fold change with respect to the contraction observed in mock-transfected WT cells. n = 3 experiments.

(C) Confocal immunofluorescence images of CAV1 and YAP in cancer-associated fibroblasts (CAFs) extracted from the stroma of human pancreatic tumors and cultured in vitro. Nuclei were detected with Hoechst (blue). Symbols mark cells with low (^∗) or high (⁺) CAV1 levels.

(D) Plot of YAP nuclear-to-cytosolic ratio against CAV1 intensity for individual CAFs as in (C). N = 97.

(E) Quantification of YAP subcellular distribution (left) and CAV1 levels (right) in CAFs transfected with CAV1 siRNAs or controls. CAV1 and YAP were detected by confocal immunofluorescence microscopy. A total of 4 independent experiments (n = 4) were analyzed (∼500 cells per experiment and condition) using Columbus. See STAR Methods for details.

Data are presented as means ± SD in (B) and means ± SEM in (E); ^∗p < 0.05, ^∗∗∗p < 0.005, and ^∗∗∗∗p < 0.0005.