Fig. 2: Select CADASIL mutant proteins have increased probability of having free thiols accessible for NEM labeling compared to WT protein.

a Demonstrates a schematic of the first 3 EGF like repeats with the predicted disulfide pairing. All cysteines examined are labeled from 1 to 18. Cysteines that are only present in mutant proteins are not shown but can be found in Fig. S1. R90C mutant protein demonstrated a significantly increased probability of being labeled with NEM at cysteine positions 4, 6, 7, and 8 (p < 0.05; (b)). The R141C mutant protein had significantly increased probabilities of being labeled with NEM at position 15 (p < 0.05) and trended toward increased NEM labeling at position 16 (c). The C49Y mutant did not significantly differ in NEM labeling probabilities from WT protein, but the mutant protein trended towards having an increased probability of NEM labeling at cysteine position 9 (d). The R75P mutant protein did not differ significantly from WT protein in NEM labeling at any cysteine position (e). The bar below each panel represents statistical significance at each cysteine position (box). Locations of each mutation found in the mutant proteins are indicated with a red arrow. Cysteines that were not detected in a sample are denoted with “nd.” Experiments were repeated at least six times using distinct samples with similar results. Figure 2 includes data from all replicates performed. Individual data points used for analysis can be found in Fig. S1b–e.