Figure 4. Effect of missense mutations on Codanin-1.

A. Pathogenic Codanin-1 mutations observed Homozygously, or compound heterozygously with a loss of function mutation (‘not LOF’), are predicted to be significantly more accessible to solvent (i.e. less buried) by the program NetSurfP-2.0 than other pathogenic mutations (‘possibly LOF’), but are not significantly different from putatively benign variants observed in the human population (gnomAD). B. Representative image of quantitative western blots using ex vivo cultured patient erythroblasts detecting Codanin-1 (upper) and C15orf41 (lower). C. Quantification of western blots shown in C showing normalized values of Codanin-1 and C15orf41 proteins compared with erythroblasts cultured from three normal individuals. Bars show the mean +/- SD. D. Box and whiskers plot showing median, quartiles and outliers of mRNA levels of CDAN1 and C15orf41 in CDA-I patients plotted relative to the levels seen in healthy controls (patient samples colour coded as in C) E. Erythroid cell counts showing that altered maturation rates do not explain the lack of protein degradation seen in this experiment. ns, not significant. * p=<0.05, **p=<0.005, significance determined by one-way ANOVA with Sidak’s multiple comparison test.