Anti futsch

Wandering third-instar larvae of both sexes from various genotypes were dissected and fixed in Bouin’s fixative for 15 min and processed for immunohistochemistry as previously described (Chen et al., 2012 (link)). Confocal images of all genotypes of larvae belonging to the same experimental group were acquired using the same settings with a Zeiss LSM710 confocal microscope and image editing was done using Adobe Photoshop. Primary antibodies used were FITC-conjugated anti-HRP (1:250, Jackson ImmunoResearch Laboratories), guinea pig anti-Ringer (1:250, Mino et al., 2016 (link)), anti-Ac-Tub (1:1000, T7451, Sigma), anti-Tubulin (1:1000, 2144S, Cell Signaling), and mouse monoclonal anti-Dlg (1:1000, 4F3), anti-Brp (1:250; NC82), anti-Futsch (1:1000; 22C10), and anti-GluR IIA (1:250, 8B4D2) were obtained from Developmental Studies Hybridoma Bank (DSHB), University of Iowa. Secondary antibodies conjugated to Alexa 488 and 568 (Invitrogen-Molecular Probes) were used at 1:400 dilution.

Wandering third-instar larvae for various genotypes were dissected and fixed in Bouin’s fix for 15 minutes and processed simultaneously as previously described^{11 (link)}. Confocal images of all genotypes of larvae belonging to the same experimental group were acquired using same settings with a Zeiss LSM710 confocal microscope.
The following primary antibodies were used: FITC-conjugated anti-HRP (1:250, Jackson ImmunoResearch laboratories), Rabbit anti-Syt^{31 (link)} (1:1000, H. Bellen, Baylor College of Medicine), mouse monoclonal anti-Dlg (1:1500, 4F3), anti-Brp (1:250, NC82), anti-DCSP2 (1:250, 6D6) and anti-Futsch (1:1000, 22C10) obtained from Developmental Studies Hybridoma Bank (DSHB), University of Iowa. Anti-Hrp conjugated to Alexa 488 or 568 were used at 1:250. Secondary antibodies conjugated to Alexa 488, 568, and 647 (Invitrogen-Molecular Probes) were used at 1:200.