Fig. 5: PREP1 affects YAP-TAZ mechanotransduction.

a Immunofluorescence images showing YAP-TAZ localization in control (siLUC) and PREP1 depleted (siPREP1) U2OS cells. Scale bar is 40 µm. Box plot shows the ratio of nuclear to cytoplasmic fluorescence intensity of YAP-TAZ in two independent experiments. N = 687 cells for siLUC and 567 cells for siPREP1. b Immunofluorescence images showing YAP-TAZ localization in control (siLUC) and PREP1 depleted (siPREP1) cells grown on soft (0.5kPa) or hard substratum (normal cell culture dishes). Scale bar is 20 μ. Box plot below shows the nuclear to cytoplasmic ratio of YAP-TAZ. N = 68 (siLUC hard), 81 (siLUC soft), 88 (siPREP1 hard) and 48 cells (siPREP1 soft). p = 0.031 (**), p = 0.0003 (***) and p < 0.0001 (****). c A representative immunoblot showing s-127 phosphorylation status of YAP-TAZ in control (siLUC) and PREP1 depleted (siPREP1) U2OS cells grown in soft (0.5kPa) vs hard (plastic) substrate. Total YAP and GAPDH act as loading controls. The data is representative of three independent experiments. Graph below shows the ratio of total YAP to phosphorylated YAP. Error values represent mean ± SE. d A representative immunoblot showing total PREP1 levels in control (siLUC), SUN1 depleted (siSUN1) or SUN2 depleted (siSUN2) U2OS or HeLa cells. GAPDH serves as loading control. Data is representative of three independent experiments. Corresponding bar graph shows densitometric analysis of band intensity from three independent experiments. Error bars represent mean ± SEM. e Bar graph shows PREP1 mRNA transcript levels in siLUC, siSUN1 or siSUN2 U2OS cells. Graph is derived from three independent experiments. Error values represent mean ± SE. f A representative immunoblot showing total PREP1 levels in control (siLUC) and LAMIN B1 depleted (siLMNB1) U2OS cells. Data is representative of two independent experiments. Bar graph shows the densitometric analysis from two experiments.