Fig. 2
Dooku1 inhibits FFSS induced Piezo1 activation and Cx43 HCs opening. A MLO-Y4 cells were pre-incubated with 40 μM Dooku1 for 30 min, then placed in the FFSS chamber, fixed on the objective table of Nikon Swept Field confocal microscope, and live images were taken at 2 s intervals for a total 15 min. Representative calcium images, first images of each phase were presented, scale bar = 20 μm; B Real-time quantification of calcium signals in Dooku1 pre-incubated and control groups, each curve represented one cell, n = 6 per group, A and B are representative images and data of three independent experiments. C EtBr dye uptake level in MLO-Y4 cells pre-incubated with 40 μM Dooku1, followed by low Ca2+ medium treatment; D EtBr dye uptake level in MLO-Y4 cells pre-incubated with different concentrations of Dooku1 (10, 20, 30, and 40 μM), followed by FFSS (16 dynes/cm2, 10 min) stimulation; E EtBr dye uptake level in MLO-Y4 cells pre-incubated with 40 μM Dooku1, followed by different magnitudes of FFSS (2, 4, 8, and 16 dynes/cm2, 10 min) treatment. F EtBr dye uptake level in MLO-Y4 cells pre-incubated with 2 μg/mL Cx43(E2) antibody, followed by FFSS (16 dynes/cm2, 10 min) stimulation. Data are shown as mean ± SEM, C–F n = 3. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Statistical analysis was performed using One-way ANOVA for multiple comparison analysis for panels C, D, and E and Two-way ANOVA for panel F