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. 2021 Oct 28;220(12):e202005184. doi: 10.1083/jcb.202005184

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© 2021 Kumari et al.

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Figure 2. — LIC1-CTD phosphorylation is required for mitotic kinetochore dynein recruitment and interaction with SAC components. (A and B) Representative confocal micrographs of nocodazole-treated prometaphase cells, immunostained for kinetochores (CREST), HC (A), and IC (B). (C and D) Quantified immunofluorescence intensity of HC (C) and IC (D) normalized to the respective CREST intensity. n = 2 independent experiments, ≥80 prometaphase cells per condition. (E and G) Immunoblots depicting SBP affinity precipitates (APs) obtained from prometaphase (E) and metaphase (G) lysates of SST-MTAP and AAA-MTAP stable cell line lysates, probed (immunoblot) with the indicated antibodies. (F and H) Quantification of AP efficiency from three independent experiments for E and G, respectively. y axes, densitometric band intensity ratios of the elute bands for the indicated proteins (x axes) normalized to the respective IC bands. Error bars, mean ± SEM over three independent AP experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (G, Kruskal–Wallis test; H, one-way ANOVA). Scale bar = 10 µm.