Immunohistochemistry was performed as described previously (12 (link)). Fluorescence images were acquired using Zeiss LSM 800 Confocal Microscope and processed with Fiji ImageJ imaging software.
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.
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.
