Fig. 2: Drpr is N-glycosylated.

a Schematic diagram showing the full-length Drpr protein with its various domains (EMI domain, green; EGF repeats, blue; and transmembrane domain, yellow) and N-glycosylation sites, as determined by MS. Blue stars denote paucimannose or high-mannose modifications and red stars denote hybrid and/or complex N-glycan modifications. b Western blot showing N-glycosylated Drpr at higher molecular-weight positions in w¹¹¹⁸ control pupal lysates, and at lower molecular-weight positions in PNGase F-treated lysates, or in lysates prepared from Alg10^wgd and xit^A homozygous pupae. c Western blot showing Drpr-HA signals in transfected S2 cells, which shift to lower molecular-weight positions upon PNGase F treatment. Drpr^NQ4-HA signal migrated to the position between those of Drpr-HA and PNGase F-treated Drpr-HA. b, c the experiments were repeated at least 3 times independently with similar results. d Schematic diagrams showing three major types of N-glycans: high mannose, hybrid and complex. Blue squares: N-Acetylglucosamine (GlcNAc), green circles: mannose, and yellow circles: galactose. e Overlaid extracted ion chromatograms of the N-glycopeptides identified by LC-MS/MS for each of the four sites (N183, N358, N504, N630) of Drpr and Drpr^NQ4. The 5 most abundant (or 3 for N630) glycoforms along with those identified as carrying HexNAc≥3 were plotted and listed in the order of signal intensities.