Dissection and staining protocol were reported previously (Deng et al., 2015 (link)). In brief, intact guts were fixed at room temperature for 45 min in 100 mM glutamic acid, 25 mM KCl, 20 mM MgSO4, 4 mM sodium phosphate, 1 mM MgCl2, 4% formaldehyde. All subsequent incubations were done in PBS, 0.5% BSA, 0.1% TritonX-100 at 4°C. rabbit anti-pH3 (Y408884, Applied Biological Materials Inc.) 1:500, rabbit anti-cleaved-caspase-3 (Asp175 Antibody, #9661, Cell Signaling Technology) 1:200. Fluorescent secondary antibodies were obtained from Jackson ImmunoResearch. DAPI was used to stain DNA.
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