Fig. 1
From: Cx43 hemichannels and panx1 channels contribute to ethanol-induced astrocyte dysfunction and damage
Ethanol increases the activity of Cx43 hemichannels and Panx1 channels in cultured astrocytes. (A) Averaged Etd uptake rate (AU/min) normalized with the control condition (dashed line) by astrocytes treated for 24 h with different concentrations of ethanol (red circles). *p < 0.05, **p < 0.005, ***p < 0.0001, ethanol treatment compared to control conditions (one-way ANOVA followed by Tukey’s post-hoc test). (B-E) Representative immunofluorescence images depicting Etd and GFAP staining from dye uptake measurements (10 min exposure to Etd) in astrocytes under control conditions (B-C) or treated for 24 h with 100 mM ethanol (D-E). (F) Averaged Etd uptake rate (AU/min) normalized with the control condition (dashed line) by astrocytes treated for several time periods with ethanol at two concentrations: 25 mM (blue circles) or 100 mM (red circles). *p < 0.05, **p < 0.005, ***p < 0.0001, 100 mM ethanol treatment compared to control conditions; #p < 0.05, ##p < 0.005, ###p < 0.0001, 25 mM ethanol treatment compared to control conditions (one-way ANOVA followed by Tukey’s post-hoc test). (G) Averaged Etd uptake rate (AU/min) normalized with control condition (dashed line) by astrocytes treated for 24 h with 100 mM ethanol alone or in combination with the following blockers: 200 µM La3+, 5 µM carbenoxolone (CBX), 500 µM Probenecid (Prob), 50 µM gap19, 50 µM gap19I130A, 50 µM 10panx1, siRNACx43, siRNAPanx1; siRNAscrb and 50 µM gap19 + 50 µM 10panx1. ***p < 0.0001, ethanol compared to control; #p < 0.05, ##p < 0.005; ###p < 0.001; effect of pharmacological agents compared to ethanol treatment (one-way ANOVA followed by Tukey’s post-hoc test). (H) Time-lapse measurements of Etd uptake by astrocytes under control conditions (white circles) or treated for 24 h with 100 mM ethanol alone (red circles) or in combination with 50 µM gap19 + 50 µM 10panx1 (black circles). Data were obtained from at least three independent experiments with three or more repeats each one (≥ 30 cells analyzed for each repeat). Calibration bar = 30 μm