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Fig. S6
Grxcr1 antibody is specific, and anti-Ush1ga and commercial anti-Ush1g antibodies recognize Ush1ga but not Ush1gb. Related to Figures 5 and 6. (A) Schematic of Grxcr1 protein showing the epitope (outlined in red) recognized by the anti-Grxcr1 antibody. (B) MDCK cells were transfected with grxcr1-HA and immuno-labelled using anti-Grxcr1 and anti-HA antibodies. Grxcr1 was detected within transfected cells as shown by the co-localization between Grxcr1 (green) and HA (red). Background at the cell membrane was also observed. (C) MDCK cells were transfected with pcDNA3.1 and immuno-labelled using anti-Grxcr1 and anti-HA antibodies. Only background at the cell membrane could be detected. Immuno-labelling using anti-Grxcr1 antibody in grxcr1+/+ (D) and grxcr1-/- mutant larvae (D’) at 5 dpf. (E) Immuno-labeling using KDEL antibody in Tg[myo6b:kdel-crimson]b1319 larvae at 5 dpf. (F) Schematic of Ush1ga protein showing the epitopes recognized by the anti-Ush1ga antibody (orange line), the mouse anti-Ush1g antibody (red line), the goat anti-Ush1g antibody (blue dotted line) and the rabbit anti-Ush1g antibody (purple line). ANK domains are indicated in green and the SAM domain is shown in pink. (G) MDCK cells were transfected with ush1ga-GFP. Ush1ga was detected within transfected cells as shown by the co-localization between Ush1ga (red) and GFP (green) signals. Background was also observed in some untransfected cells. (H) MDCK cells were transfected with ush1gb-HA. Ush1ga (red) could only be detected at background level in transfected cells (green). (I) MDCK cells were transfected with ush1ga-GFP and immuno-labelled with rabbit anti-Ush1g and anti-GFP antibodies. Ush1ga was detected in transfected cells as shown by the co-localization between Ush1g (red) and GFP (green) signals. (J) MDCK cells were transfected with ush1gb-HA and immuno-labelled with rabbit anti-Ush1g and anti-GFP antibodies. Ush1gb could not be detected in transfected cells. (K) MDCK cells were transfected with ush1ga-GFP and immuno-labelled with goat and mouse anti-Ush1g antibodies and anti-GFP antibody. Ush1ga was detected in transfected cells as shown by the co-localization between Ush1g (red-goat and blue-mouse) and GFP (green) signals. (L) MDCK cells were transfected with ush1gb-HA and immuno-labelled with goat and mouse anti-Ush1g antibodies and anti-GFP antibody. Ush1gb could not be detected in transfected cells.