Ab83528

The antibodies and their concentrations used for Western blotting are as follows: anti‐mAcon1 mouse (1 µg/ml; ab83528, Abcam), anti‐ATG8A rabbit (1:1000; ab109364, Abcam), anti‐β‐actin rabbit (1:1000; 4967S, Cell Signaling), anti‐mouse goat IgG‐horseradish peroxidase (HRP) (1:10000; sc2005, Santa Cruz Biotechnology), and anti‐rabbit goat IgG‐HRP (1:10000; sc2004, Santa Cruz Biotechnology) antibodies. The antibodies and their concentrations used for IHC are as follows: anti‐mAcon1 mouse (1:100; ab83528, Abcam), anti‐BRP mouse (1:25; DSHB), anti‐ATG8A rabbit (1:200; ab109364, Abcam), anti‐GFP rabbit (1:1000; Invitrogen), anti‐GFP mouse (1:500; GFP‐G1, DSHB), anti‐mouse goat (1:1000; Alexa Fluor^® 488, Invitrogen), anti‐rabbit goat (1:1000; Dylight^® 488, Invitrogen), and anti‐rabbit goat (1:1000; Alexa Fluor^® 594, Invitrogen) antibodies.

Intact guts were fixed at room temperature for 20 min in PBS, 3.7% formaldehyde. All subsequent incubations were done in PBS, 4% horse serum, 0.2% Triton X-100 at 4°C following standard protocols. To visualize the three-dimensional arrangement of the internal organs inside the male body cavity, intact abdomens were fixed at room temperature for 20 min in PBS, 3.7% formaldehyde prior dissection and cuticle removal.
The following primary antibodies were used: chicken anti-GFP (ab13970, Abcam) 1/10000, mouse anti-GFP (11814460001, Roche) 1/1000, chicken anti-beta Galactosidase (ab9361, Abcam) 1/200, rabbit anti-phospho-Histone H3 Ser10 (9701L, Cell Signaling Technology) 1/500, mouse anti-Fas3 (7G10, DSHB) 1/50, rabbit anti-Pyruvate dehydrogenase E1-alpha subunit (phospho S293) (ab92696, Abcam) 1/200, rabbit anti-Aconitase 2 (ab83528, Abcam) 1/200, rabbit anti-cleaved Drosophila Dcp-1 (Asp216) (9578S, Ozyme) 1/500, and rhodamine Phalloidin (R415, ThermoFisher scientific) 1/1000. Fluorescent secondary antibodies (FITC-, Cy3- and Cy5-conjugated) were obtained from Jackson Immunoresearch. Vectashield with DAPI (Vector Labs) was used to stain DNA.