Figure S3.

In silico prediction of transmembrane helices and N-glycosylation sites in huCLN3, zfcln3, and CRISPR mutants.

The TMHMM (v2.0) program predicts 11 putative transmembrane (helical) regions in the wild-type human and zebrafish CLN3 proteins (73Preprint). In contrast, only one transmembrane domain is predicted for the truncated protein expressed by the MUT1 line, whereas the protein encoded by the cln3^Δex4 allele (MUT2 line) is predicted to have a shorter luminal loop between the first two transmembrane domains. The NetNGlyc (v1.0) tool predicts four glycosylation sites in human CLN3 (74). N49 is inside of a transmembrane domain and therefore unlikely to be glycosylated. In contrast, the zebrafish protein contains five predicted N-glycosylation sites outside of the transmembrane domains. The expression product of the MUT1 allele is predicted to conserve the first four glycosylation sites, whereas the one of the cln3^Δex4 allele conserves only the first of these glycosylation sites that is placed in exon 3.