Fig. 4

The expression and distribution of Piezo1 in MDA-MB-231 cells were regulated by caveolae. a Representative fluorescence images of Piezo1 (magenta) and caveolae (green) colocalization visualized by confocal microscopy (100X) and 2D intensity histogram output in MDA-MB-231 cells. Insets in both conditions show a magnified view of the boxed regions. b Representative image of 2D intensity histogram output of Coloc2 analysis performed using Fiji software. The text indicates the Pearson coefficient of the pixel-intensity correlation (n = 8). c Western blot images and quantification of Piezo1 expression in wild type (WT), Cav-1 EGFP expressing, and Cav-1 KD MDA-MB-231 cells (means ± s.e.m., n = 3). Cropped images of Western blots are shown and uncropped images are shown in Fig. S8b. ^** p < 0.01 versus WT groups. d, Representative fluorescence images of Piezo1 (green) and nucleus (blue) visualized by confocal microscopy (100X) after cells were treated with MβCD for 5 min, 10 min, and 20 min (upper panel: x-y view, lower panel: x-z view, white dashed line shows the position of a section of x-z view). e Time-courses of relative mean fluorescence intensity of G-GECO in MDA-MB-231 cells pretreated with or without MβCD, and Cav-1 KD in response to 400 Pa compression. Each experiment assayed 10–20 cells and repeated three times. The black bar indicates the period of compression. f Quantification of the fold change of invaded cells treated with siRNA for Cav-1 under 400 Pa. Data are presented as means ± s.e.m., n = 3, ^** p < 0.01 versus Ctr groups