Fig. 5
Missense variants alter the maturation and cell-surface levels of SorLA protein in hiPSCs. a Western blot analyses of glycosidase treatments of wild-type full-length SorLA (SorLAFL) expressed in hiPSC at endogenous level. Cellular lysates were treated in the absence or presence of the glycosidases PNGase F, Neuraminidase (Neu), O-Gycosidase (O-Glyc). Untreated samples (–) were incubated under the same conditions but without enzymes. Maturation profile of wild-type and SorLA missense variants expressed in hiPSC at endogenous level. c Surface biotinylation experiments examining the cell-surface steady-state level of SorLA protein in wild-type or S124R, R332W, N371T, S577P and R654W CRISPR/Cas9-edited hiPSC. The cytosolic FUS protein was used as control to exclude non-specific labelling of the intracellular fractions. Several independent clones (#) are presented. c Blots were probed with an anti-SorLA antibody and representative blots are presented. Immature core-glycosylated and mature complex-glycosylated SorLA are indicated with solid and empty arrowheads, respectively. The molecular masses of marker proteins in kDa are shown on the left. d Normalized quantification from at least 2 independent experiments. WT mean was arbitrarily set at 100 units. Protein levels were compared by using a regression model adjusted on the experiment as random effect. P-value significances are displayed after Bonferroni’s correction for 5 statistical tests