The primary antibodies used were as follows: chicken anti-GFP (1:1,000) (ab13970; Abcam), rabbit anti-HA (1:1,000) (3724S; Cell Signaling Technologies), rabbit anti-DsRed (1:200) (632496; Takara), and mouse anti-chaoptin (1:20) (24B10; Developmental Studies Hybridoma Bank). Rat anti-dFezf (1:50) was a gift from Dr. Claude Desplan (New York University, New York, NY).
The secondary antibodies used were as follows: Goat anti-chicken immune globulin (Ig) G (H + L) Alexa Fluor 488 (A-11039), goat anti-rabbit IgG (H + L) Alexa Fluor 568 (A-11011), goat anti-mouse IgG (H + L) Alexa Fluor 647 (A-21236), and goat anti-rat IgG (H + L) Alexa Fluor 568 (A-11077) were purchased from Thermo Fischer Scientific and used at a dilution of 1:500.