(A) Location of acidic and basic residues within crystal packing of ISB between INCENP1 and Borealin2/Borealin3. Side chains are colored in red to indicate acidic charge and blue to indicate basic charge. (B) Summary of a second round of mutations made to acidic residues within INCENP. Lolli-pop sticks represent each of the indicated residues in question. (C) DIC micrographs of the ISB droplets for IMut3SB, IMut4SB, and IMut5SB. Scale bar = 10 µm. (D) Turbidity calculations of IMut3SB, IMut4SB, and IMut5SB measured as absorbance at 330 nm; n=6 for (ISB)WT, IMut4SB, and IMut5SB. n=3 for IMut3SB. Statistical analysis was performed using a Brown-Forsythe and Welch ANOVA test. ****p<0.0001; **0.001<p<0.01. (E) Summary of a third round of mutations made to acidic residues within INCENP. Lolli-pop sticks represent each of the indicated residues in question. (F) DIC micrographs of the ISB droplets for IMut6SB and IMut7SB. Scale bar = 10 µm. (G) Turbidity calculations of IMut6SB and IMut7SB measured as absorbance at 330 nm; n=6 for (ISB)WT. n=3 for IMut6SB and IMut7SB. Statistical analysis was performed using a Brown-Forsythe and Welch ANOVA test. ****p<0.0001; *0.01<p<0.05.
Figure 4—source data 1. Data used to generate Figure 4.