Fig. 3: Variants Increasing Insulin Binding.

A Scatter plot of median cell surface expression scores against insulin binding scores, focused only on gain of insulin binding (score > 0). B Similar scatter plot of individual missense variants with significantly increased insulin binding scores (score > 0 and FDR < 0.01). Variants at G333 and D627 are shown in red and blue respectively. C Binding of increasing concentrations of Eu-labelled Insulin to WT and mutant INSR over 16 hr at 4 °C. Data from 3 replicates are shown, represented as mean ± standard deviation (D) Representative immunoblot of lysates of MEFs expressing either WT or mutant INSR and stimulated for 10 min at 37 °C with increasing concentrations of insulin. E Densitometric analysis of Western blot images from 3 independent experiments, represented as mean ± standard deviation. F Variant effect map excerpt with highlighted gain-of-insulin-binding hotspots, visually ascertained.