Rabbit anti cleaved caspase 3 d175

Wandering third instar larvae of mixed sex were gross dissected in PBS using sharpened forceps to expose the larval brain. The inverted carcasses were fixed in 4% formaldehyde, washed three times in PBT (0.1% Triton X-100 in PBS), blocked in PBT-G (1% normal goat serum+0.3% Triton X-100 in PBS) and incubated in primary antibody diluted in PBT-G overnight at 4°. The next day, the inverted carcasses were washed three times in PBT, incubated in secondary antibody+Phalloidin-FITC for one hour at 25°, and washed three additional times in PBS before fine dissecting the larval brains. The brains were mounted on a slide with mounting media containing DAPI. The slides were imaged using a Zeiss 710 confocal microscope, and the images were processed with ImageJ (https://imagej.nih.gov/ij/). The following antibodies were used in this study: rabbit anti-cleaved caspase 3 D175 at 1:100 (Cell Signaling Technology #9661); Alexa 647 goat anti-HRP at 1:500 (Jackson ImmunoResearch Laboratories Inc. 123-605-021); Alexa 488 goat anti-rabbit at 1:1000 (Life Technologies ref# A11008).

The following primary antibodies were used for immunohistochemistry (IHC), immunofluorescence (IF) and western blots (WB): rabbit anti-Vangl2 (Aviva Systems Biology, San Diego, CA, OAAB15535; 1:200, IHC), rabbit anti-Vangl2 (Ramsbottom et al., 2014 (link)) (21st Century Biochemicals, Marlborough, MA; 1:2000, WB), rabbit anti-Gapdh (Millpore, Temecula, CA, ABS16; 1:30,000, WB), rabbit anti-cofilin (Cell Signaling, Beverly, MA, 5175; 1:1000, WB), rabbit anti-p-cofilin (Ser3) (Cell Signaling, 3313; 1:1000, WB), rabbit anti-β-catenin (Cell Signaling, 9562; 1:1000, WB), mouse anti-β-actin (MP Biomedicals, 08691001; 1:10,000, WB), mouse anti-E-cadherin (BD Biosciences, 610181; 1:2500, WB), rabbit anti-GM130 (Sigma-Aldrich, G7295; 1:1500, IF), rabbit anti-elastin (Biorbyt, orb13391; 1:100, WB), rabbit anti-MMP-12 (Abcam, Ab52897; 1:1000, WB and 1:200, IHC), Armenian hamster anti-Pecam1 (Abcam, Ab119341; 1:500, IF; a gift from Leo Carlin), rabbit anti-phospho-histone H3 (Ser10) (PH3; Millipore, 06-570; 1:1000, IF), rabbit anti-cleaved caspase 3 (D175) (Cell Signaling, 9661; 1:1000, IHC), goat anti-CC10 (T-18) (Santa Cruz Biotechnology, Dallas, TX, sc-9772; 1:1000, IHC), rabbit anti-proSP-C (Millipore, AB3786; 1:1000, IHC), rabbit anti-non-phospho (active) β-catenin (Cell Signaling, 8814; 1:1000, WB).