Extended Data Figure 6: Protein expression of uncoupled mutations.

In order to determine which family 1 DCHS1 mutation is leading to the observed decrease in protein expression, constructs were generated that harboured only the p.P197L or the p.R2513H variant. Mutant refers to the double mutant P197L/R2513H construct. Western blot analyses from transfected HEK293 cells, three independent biological replicates, demonstrate that the p.R2513H mutation causes a significant decrease in DCHS1 protein expression, similar to that of the construct with both variants (mutant), suggesting pathogenicity. Percent difference in protein levels is depicted. Normalization of data was accomplished by qPCR specific to the transfected constructs. P values from the Student’s t-test are indicated in graphs.