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

Figure 5.

Figure 5

Acetaldehyde modulates SMAD7 expression. A: Total RNA was obtained from HHSCs cultured in the absence or presence of 200 μmol/L acetaldehyde for times ranging from 30 minutes to 48 hours. Acetaldehyde down-regulates the expression of SMAD7 mRNA in a time-dependent manner, reaching the maximum down-regulation of 60% after 3 hours, relative to the value at 30 minutes, after which time expression increases steadily and reaches values threefold above basal levels by 48 hours. SMAD7 mRNA expression levels were corrected for differences in loading using GAPDH as a housekeeping gene. ∗P < 0.05 versus control. B: HHSCs were transiently cotransfected with the −378COL1A2 reporter plasmid and a cytomegalovirus-driven SMAD7 expression vector. At 12 hours after transfection, cells were treated with 200 μmol/L acetaldehyde for 36 hours. Cells were harvested and used to measure luciferase activity. Acetaldehyde failed to up-regulate the expression of COL1A2 reporter vector in HHSCs cotransfected with a vector overexpressing SMAD7 and the −378COL1A2LUC reporter vector. Data are expressed as means ± SEM. ∗P < 0.05 versus acetaldehyde-treated empty vector. AU, arbitrary units.