Hh signaling is required for DSS-stimulated Upd2 expression. (A–B′) Expression of 10XStat-dGFP in adult midguts expressing Su(H)^ts (Con; A and A′) or Su(H)^ts>Smo^RNAi (B and B′) and treated with Suc or DSS for 1 d. (C–F) Adult female flies expressing esg^ts or esg^ts>Stat^RNAi for 8 d at 29°C were treated with Suc or DSS for 1 d before midguts were dissected out and immunostained for PH3 and DRAQ5. (G) Quantification of PH3⁺ cells in midguts of the indicated genotypes treated with Suc or DSS after adult flies were raised for 10 d at 29°C. Three independent experiments were performed and 20 guts were examined for each sample. Error bars are standard deviations. **, P < 0.01. (H) Quantification of Upd2 mRNA levels by RT-qPCR in Su(H)^ts or Su(H)^ts>Smo^RNAi midguts treated with Suc or DSS. Three independent experiments were performed and error bars are standard deviations. **, P < 0.01. (I–L) Adult flies expressing Su(H)^ts or Su(H)^ts>Upd2^RNAi for 10 d at 29°C were treated with Suc or DSS for 1 d before midguts were dissected out and immunostained for PH3 and DRAQ5. (M) Quantification of PH3⁺ cells in midguts of the indicated genotypes treated with Suc or DSS. Three independent experiments were performed and 20 guts were examined for each sample. Error bars are standard deviations. **, P < 0.01. Bars, 50 µm.