Fig. 3
SorLA variants associated with abnormal immunoblot profiles interfere with the secretion of SorLA truncated constructs in HEK293 cells. a Schematic representation of the soluble truncated SorLA759 and SorLA2131 proteins, and the location of SorLA variants examined. The Mis3, Mis2, and Mis0-1 missense variants studied are shown in red, orange and black, respectively. Maturation-deficient SorLA variants are underlined. b Wild-type (WT) and mutants SorLA proteins were expressed in HEK293 cells. The SorLA proteins secreted into the cellular medium were analyzed by immunoblotting using an anti-SorLA antibody. Total protein was used as the loading control with Stain-free (SF) technology. Representative blots are presented. Maturation-deficient SorLA variants are underlined. The molecular masses of marker proteins in kDa are shown on the left. c Quantification of secretion efficiency from 4 independent experiments (SorLA759) or 3 independent experiments (SorLA2131). The normalized quantification of the SorLA proteins in the cellular medium is reported in the graphs. For both SorLA759 and SorLA2131, WT-SorLA were arbitrarily set at 100 arbitrary units. Protein levels were compared by using a regression model adjusted for experiment as random effect. P-value significances are displayed after Bonferroni’s correction for 14 and 17 statistical tests for SorLA759 and SorLA2131, respectively