Fig. 5

Fig. 5 Cholesterol depletion activates Latrophilin-dependent signaling. (A) ECs were treated with FSS or 5 mM methyl-β-cyclodextrin (mβCD) for 1 min then incubated with D4H-mClover3, rinsed and bound mClover3 quantified as described in “Methods”. N = 27 for each condition. ****P < 0.0001; one-way ANOVA with Tukey multiple comparisons test. (B) Phalloidin staining following short acute treatment of methyl-β-cyclodextrin (mβCD). Scale bar: 100 µm. (C) Co-immunoprecipitation of Gi1Q306K(ins4A) with endogenous Latrophilin-2 (LPHN2) after 5 mM MβCD for 1 min. (D) Activation of Src family kinases and VEGFR2 after 5 mM MβCD for 1 min in HUVECs ± LPHN2 knockdown. Results quantified in (E, F), respectively. Values are means ± SEM. N = 5. *P = 0.0305, ***P = 0.0002; one-way ANOVA with Tukey multiple comparison test. (G) Model for junctional endothelial shear stress mechanotransduction.