Dissection, fixation and immunostaining were performed as described by [68 (link)]. The following antibodies were used: mouse anti-Prospero (MR1A-c, Developmental Studies Hybridoma Bank (DSHB)) at 1:200; mouse anti-Arm (N2 7A1-s, DSHB) at 1:50; rabbit anti-Caspase3 (Cell Signaling, #9661) at 1:300; rabbit anti-DH31 (gift from Jan Veenstra [40 (link)]) and Michael Nitabach [31 (link)]) at 1:500. The secondary antibodies used were anti-mouse Alexa647, anti-rabbit Alexa488, anti-rabbit Alexa546 (Invitrogen). All secondary antibodies were used at 1:1000. Phalloidin-Alexa555 (Molecular Probes, A34055) were used at 1:500 2h at room temperature or 1:2000 overnight at 4°C. Guts were mounted in Fluoroshield-DAPI medium (Sigma) and observed with a Zeiss Axioplan Z1 with Apotome 2 microscope. Pictures in Fig 4M and 4N were acquired using a Zeiss LSM 880 confocal equipped with a Fast AiryScan. Images were analyzed using ZEN (Zeiss) and Photoshop software. Image acquisition was performed at the Microscopy platform of the Institut Sophia Agrobiotech (INRA 1355-UNS-CNRS 7254-Sophia Antipolis).
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