Hh signaling is required in precursor cells for injury-induced ISC proliferation and midgut regeneration. (A–I) 2- to 3-d-old adult females of esg^ts without (A–C) or with UAS-Smo^RNAi (D–F) or UAS-Ci^RNAi (G–I) were shifted to 29°C for 8 d and then treated with Suc, DSS, and bleomycin for 1 d, followed by immunostaining for GFP and PH3. (J) Quantification of PH3⁺ cells in midguts from adults of the indicated genotypes. Three independent experiments were performed and 12 guts were examined for each sample. Error bars are standard deviations. **, P < 0.01. (K–R) 3- to 5-d-old adult females of esg^F/O without (K–N) or with UAS-Smo^RNAi (O–R) were shifted to 29°C for 8 d and treated with Suc or DSS for 1 d, followed by immunostaining for GFP and LacZ (K, L, O, and P) or GFP, Pdm1, and Pros (M, N, Q, and R). (S) Quantification of GFP⁺ or Su(H)-lacZ⁺ cells in midguts of the indicated genotypes treated with Suc or DSS for 1 d. n = 20 for each genotype. Error bars are standard deviations. **, P < 0.01. Bars, 50 µm.