Fig. 3

Ethanol does not modulate levels and plasma membrane distribution of Cx43 and Panx1 in cultured astrocytes. (A-L) Representative confocal images depicting Cx43 (cyan, left panel) or Panx1 (cyan, right panel) immunostaining in combination with DAPI (magenta) and WGA (red) labeling by astrocytes under control conditions (A-C and G-I) or treated for 24 h with 100 mM ethanol (D-F and J-L). The size of the z-step acquisition for all images was 0.3 μm. Calibration Bar: 10 μm. Insets: 2X magnification of the indicated area of panels C, F, I and L. (M-N) Quantification of membrane, intracellular and total staining of Cx43 (M) and Panx1 (N) normalized to control conditions (dashed line) by astrocytes treated for 24 h with 100 mM ethanol. (O) Quantification of Manders’ overlap coefficient for Cx43 or Panx1 with WGA by astrocytes under control conditions (white bars) or treated for 24 h with 100 mM ethanol (red bars). (P) Total Cx43 (upper panel) and Panx1 (bottom panel) levels by astrocytes under control conditions or treated for 1, 24, 48–72 h with 100 mM ethanol. Total levels of each analyzed band were normalized according to the levels of GADPH detected in each lane. (Q-R) Quantification of total levels of Cx43 (Q) and Panx1 (R) normalized to control (dashed line) in astrocytes treated for 1, 24, 48–72 h with 100 mM ethanol. Averaged data were obtained from three independent experiments