Rabbit anti c myc a 14

Larvae and prepupae (0–10 h APF) were dissected in 1×PBS for isolating wing discs, which were then fixed in 4% paraformaldehyde for 15 minutes at room temperature (RT). For pupae 12–36 h APF, pupal cases were removed in 1×PBS and the whole pupae were fixed in 4% paraformaldehyde at RT for one hour. Internal pupal membranes on the surface of wing tissues were subsequently removed and the resulting wing discs were fixed again in 4% paraformaldehyde for 15 minutes. The following primary antibodies were used: mouse anti-Ac (DSHB, 1∶5) [79] (link), mouse anti-ß-galactosidase 40-1a (DSHB, 1∶500) [80] (link), mouse anti-BR-Z1 Z1.3C11.OA1 (DSHB, 1∶250) [48] (link), mouse anti-EcRA 15G1a (DSHB, 1∶100) [81] (link), mouse anti-Elav 9F8A9 (DSHB, 1∶500) [82] (link), mouse anti-Futsch 22C10 (DSHB, 1∶500) [83] (link), mouse anti-Hnt 1G9 (DSHB, 1∶25) [84] (link), mouse anti-Pros MR1A (DSHB, 1∶100) [85] (link), rabbit anti-c-Myc A-14 (Santa Cruz, 1∶500), mouse anti-Pol II CTD4H8 (Santa Cruz, 1∶500), rabbit anti-Histone H3 GTX122148 (GeneTex, 1∶500), rabbit anti-JAB1 (CSN5) (Sigma, 1∶250), and guinea pig anti-Sens (1∶2000) [3] (link). The anti-JAB1/CSN5 antibody was pre-cleaned by incubation of 10× volume antibody with CSN5^null larval brain tissues at 4°C overnight.

The following primary antibodies were used: rabbit anti-Rab20 (11616-1-AP, Proteintech, Chicago, IL); rabbit anti-Rab20 (GTX119559, Genetex, Irvine, CA); rabbit anti-Rab5a (C8B1, Cell Signaling Technology, Beverly, MA); rabbit anti-Rabex-5 (Sigma-Aldrich, Germany); mouse anti-β-actin (mAb8226, Abcam, Cambridge, UK); mouse anti-GFP (JL-8, Clontech, Mountain View, CA) and rabbit anti-c-Myc (A-14, Santa Cruz Biotechnology, Dallas, TX). Secondary antibodies conjugated to Alexa Fluor 488, 546 or 633 for indirect immunofluorescence studies were purchased from Molecular Probes (Invitrogen, Carlsbad, CA). Secondary antibodies conjugated to Cy3 or horseradish peroxidase (HRP) were obtained from Jackson ImmunoResearch Laboratories (West Grove, PA).