Figure 2.

Targeted deacetylation of the slo promoter can interfere with alcohol tolerance. (A) Schematic depiction of the genetic cross used to inducibly tether the RPD3 HDAC activity within the slo transcriptional control region: transgenic flies that carry a heat-inducible Gal4^DBD:Rpd3 cDNA (I) were crossed to flies carrying either the slo^UAS-6b-L allele (II) or to flies carrying the slo^6b-L allele that lacks the UAS site (III). (B,E) Paradigm to test for an effect of the tethered RPD3 on alcohol tolerance. Animals were sedated with ethanol (B) or benzyl alcohol (E) once (top line) or twice (bottom line; 24 h between sedations). All animals were heat treated to activate Gal4^DBD:Rpd3 expression. The time of heat activation was 30 min after the first alcohol treatment. (C,F) Average recovery time of animals after a single (black bar) or after two consecutive (gray bar) sedations with benzyl alcohol (C) or ethanol (F). See Supplementary Figure S2 for the corresponding recovery curves. Error bars represent SEM (Student’s t-test: *** in C denotes P < 0.0001, n = 32 [2nd exp], 30 [1st exp.]; ** in F denotes P = 0.0026, n = 54 [2nd exp], 43 [1st exp.]; *** in F denotes P < 0.0001, n = 47 [2nd exp], 47 [1st exp.];). (D,G) Magnitude of tolerance induced in the Hsp-Gal4^DBD:Rpd3/FM6 ; ; slo^6b-L animals (6b-L) and in the Hsp-Gal4^DBD:Rpd3/FM6 ; ; slo^UAS-6b-L (UAS-6b-L) animals. Error bars represent SEM (Student’s t-test: * in D denotes P = 0.0238, n = 32 [6b-L], 33 [UAS-6b-L]; * in G denotes P < 0.0367, n = 47 [6b-L], 43 [UAS-6b-L]).