Rat α ncad

Samples were fixed with 4% paraformaldehyde, washed with phosphate buffer with 0.2% Triton X-100, and staining as previously described. Primary antibodies were used in the following dilutions: rabbit α-GFP 1:1000 (Invitrogen), rat α-Ncad 1:20 (Developmental Studies Hybridoma Bank), mouse α-Bruchpilot 1:50 (Developmental Studies Hybridoma Bank), mouse α-rat CD2 1:200 (Serotec), rabbit α-RFP 1:200, chicken α-GFP 1:700. The following secondary antibodies were used: AlexaFluor488 goat α-rabbit 1:1000, goat α-mouse-Cy3 1:100, AlexaFluor568 goat α-mouse IgG highly cross-adsorbed 1:300, AlexaFluor647 goat α-rat 1:200, AlexaFluor633 goat α-mouse 1:200, goat α-rabbit Cy3 1:200, AlexaFluor488 goat α-rat 1:200, AlexaFluor488 goat α-chicken 1:700, goat α-rat Cy3 1:200. Confocal images were taken by an Olympus Fluoview FV1000 or Zeiss LSM 510 (Light Microscopy Core Facility).

Staining of adult and pupal brains was performed as previously described [18 (link),24 (link)]. Primary antibodies were used in the following dilutions: rabbit α-GFP 1:1,000 (Invitrogen), rat α-Ncad 1:20, rat α-elav 1:100, mouse α-Bruchpilot 1:20 (Developmental Studies Hybridoma Bank), mouse α-CD2 1:1,000 (Serotec). The following secondary antibodies were used: goat α-rabbit-FITC 1:1000, goat α-mouse-Cy3 1:100, goat α-rat-Cy3 1:200, goat α-guinea pig-Cy3 1:500, goat α-rat-Cy5 1:200, goat α-rabbit-Cy5 1:500 (Jackson ImmunoResearch), Alexa 568 goat α-mouse IgG highly cross-adsorbed 1:300 (Invitrogen). Confocal images were taken using an Olympus Fluoview FV1000.

Samples were fixed with 4% paraformaldehyde, washed with phosphate buffer with 0.2% Triton X-100, and staining as previously described [87 (link)]. Primary antibodies were used in the following dilutions: rabbit α-GFP 1:1000 (Invitrogen), chicken α-GFP 1:700 (Aves Labs), rat α-Ncad 1:20 (Developmental Studies Hybridoma Bank), mouse α-Bruchpilot 1:20 (Developmental Studies Hybridoma Bank), mouse α-CD2 1:1000 (Serotec), mouse α-Dac 2–3 1:20 (Developmental Studies Hybridoma Bank), rabbit α-β galactosidase 1:800 (Invitrogen), mouse α-β galactosidase 1:800 (Promega), rat α-Bab2 1:1500 (Frank Laski), rabbit α-Bar-H1 1:100 (Tetsuya Kojima). The following secondary antibodies were used: Alexa 488 goat α-rabbit 1:1000, Alexa 488 goat α-chicken 1:1000, goat α-mouse-Cy3 1:100, goat α-rat-Cy3 1:200, goat α-rabbit-Cy3 1:200, Alexa 568 goat α-mouse IgG highly cross-adsorbed 1:300, Alexa 647 goat α-rat 1:200, Alexa 633 goat α-mouse 1:200, Alexa 647 goat α-mouse 1:200. Confocal images were taken by an Olympus Fluoview FV1000 or Zeiss LSM 510 (Light Microscopy Core Facility).