Cell cycle dynamics & EdU Index (Fig. 5B & Fig S7C)

Juveniles were dissociated in pools of 15 polyps. Prior dissociation were labeled for 30 min with EdU (100uM, 2%DMSO) over several time points after feeding, and then dissociated and fixed using Trypsin/formaldehyde. Cells were stained with 1μg/ml FxCycle violet DNA dye (Invitrogen), EdU was labelled with Alexa647 using a Click-it reaction, and mOr2 was detected with anti-dsRed (Rabbit, Takara Bio Clontech 632496) and goat-anti-rabbit Alexa568.

Flow cytometry was performed a BD LSRFortessa (BD Life Sciences) and the mOr2 and EdU positive cells in samples were determined based on Alexa568 and Alexa647 fluorescence in comparison with the negative controls on cells within the cell cycle (based on DNA staining) using FlowJoV10.8 (BD Life Sciences).

.wsp files contain the samples, gating strategy and cell populations used for the analysis in FlowJo

.fcs files represent the individual flow cytometry output files that were analysed in the .wsp file