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. 2014 May;184(5):1458–1467. doi: 10.1016/j.ajpath.2014.01.020

Figure 9.

Figure 9

Lactacystin, an inhibitor of proteasomal degradation, inhibits acetaldehyde-mediated expression of COL1A2 promoter-driven reporter activity. A: Western analysis of Ski in nuclear proteins extracted from HHSCs treated with 200 μmol/L acetaldehyde, 30 μmol/L lactacystin, or both. In HHSCs treated with lactacystin alone, Ski levels were higher than in control cells. B: HHSCs were transfected with the −378COL1A2LUC reporter vector; 12 hours later, cells were incubated with 30 μmol/L lactacystin. After 30 minutes, acetaldehyde (200 μmol/L final concentration) was added; 36 hours later, cells were harvested and luciferase activity was determined. Controls for these experiments were transfected cells without treatment, cells treated with lactacystin alone, and cells treated with acetaldehyde alone. Lactacystin prevented acetaldehyde-mediated decrease in nuclear Ski by approximately 50% and up-regulation of the −378COL1A2LUC reporter vector activity by 50%. Data are expressed as means ± SEM. ∗P < 0.05 versus control. †P < 0.05 versus lactacystin treated cells. ‡P < 0.05 versus acetaldehyde-treated cells.