Figure 3. Daily moderate EtOH consumption reduces the number of Sca1⁺ cells in ligated carotids.

Partial carotid ligation or sham-operation was performed in Sca1-eGFP transgenic mice treated daily with or without ‘moderate’ amounts of EtOH (0.8 g/kg, oral gavage). Control ligated animals received an isocaloric cornstarch solution. Vessels harvested on day 14 were fixed, sectioned and imaged for eGFP, indicative of Sca1 expression. Daily moderate EtOH inhibited adventitial and intimal formation concomitant with a reduction in the number of Sca1⁺ cells per section as analyzed by Fiji ImageJ software. (a) Representative images (×20 magnification, scale bar 50 μM.); Verhoeff-Van Gieson stained sections (top) and corresponding confocal immunofluorescence pics (bottom) for sham, ligated, and ligated + EtOH groups. Thickness of Adventitia: ‘A’ yellow line, Media: ‘M’ white line, Neo-intima: ‘NI’ red line. (b) Higher magnification view (×60) of sham-operated carotid with a Sca1⁺ cells present in adventitia and intimal layer indicated by arrows. (c) Bar graph shows cumulative data for Sca1⁺ cells as a percentage of total cells (determined by DAPI); Sham n=5, Ligated n=6, Ligated + EtOH, n=6, *p<0.05 vs sham, #p<0.05 vs ligated).