Figure 4. Interaction of core BBSome with SSTR3 peptides.

Immobilized 15-residue peptides of SSTR3 were incubated with Flag antibody alone (A) or with the core BBSome and subsequently with Flag-antibody (B). The Flag-antibody detected either the Flag-tag on the BBS8 subunit or the Flag control peptides located at the corners of the arrays. (C) Averaged and normalized intensity of the Flag-signal from (B). The positioning of the two strongest peptide hits is presented within a schematic SSTR3 plot (D).

Figure 4—figure supplement 1. Peptide microarray with core BBSome.

15-residue peptide fragments of different potential BBSome-binding proteins were immobilized on a microarray in duplicate and analyzed for their affinity to the core BBSome. The captured BBSome was detected by a fluorescent Flag-antibody (labelled with Dylight800, PepPerPrint, Germany) that was targeted to the Flag-tagged BBS8 subunit. As a control, the microarray was first incubated with Flag-antibody alone and the corresponding fluorescence image was used to identify any direct interactions of the antibody with immobilized peptides (yellow boxes indicate these potentially false-positive signals). Relevant BBSome interactions with the third intracellular loops of GPCRs are marked as blue boxes, while interactions with the C-terminal cytoplasmic domains are marked with green boxes.