Knockdown of HDAC1-3 prevents decreases in α1 expression caused by EtOH exposure. (A) Schematic demonstrating the experimental outline for knockdown of HDAC1 and HDAC2 using shRNA delivered via lentiviral transduction. (B) Schematic demonstration of the experimental outline for knockdown of HDAC3 using siRNA via transfection. (C) Quantification of HDAC1 expression after HDAC1 knockdown (effect of HDAC1 knockdown, two-way ANOVA, F1,20 = 105.1, P < 0.0001, n = 6). (D) Quantification of HDAC2 expression after HDAC2 knockdown (effect of HDAC2 knockdown, two-way ANOVA, F1,19 = 10.24, P = 0.0047, n = 5–6). (E) Quantification of HDAC3 expression after HDAC3 knockdown (effect of HDAC3 knockdown, two-way ANOVA, F1,20 = 47.41, P < 0.0001, n = 6). (F) Representative Western blots and quantification of α1 expression after HDAC1 knockdown (effect of HDAC1 knockdown, two-way ANOVA, F1,20 = 20.84, P = 0.0002. n = 6). (G) Representative Western blots and quantification of α1 expression after HDAC2 knockdown (two-way ANOVA interaction, F1,17 = 7.521, P = 0.0139, n = 4–6). (H) Representative Western blots and quantification of α1 expression after HDAC3 knockdown (two-way ANOVA interaction, F1,19 = 9.403, P = 0.0064, n = 5–6). Each bar represents the mean ± S.E.M. Statistical significance is determined by two-way ANOVA followed by Bonferroni post hoc tests. *P < 0.05; **P < 0.01. KC, knockdown of HDAC + H2O exposure; KE, knockdown of HDAC + EtOH exposure; SC, scrambled virus or siRNA + H2O exposure; SE, scrambled virus or siRNA + EtOH exposure.