Fig. 5
From: Internal modifications in the CENP-A nucleosome modulate centromeric dynamics
K124A/Q have altered affinity for CENP-C. a Anti-HA ChIP was performed on medium-sized chromatin arrays (see Additional file 6: Fig. S4D), and the amount of co-IP’ed CENP-C (CpC) was determined. Arrow indicates location of HA-tagged CpA/K124A/K124Q versus native CENP-A (nCENP-A) below. Three independent replicates conducted using HA-tagged CpA/K124A/K124Q mutants were analyzed by measuring the ratio of CENP-C/ChIP’ed HA-tagged mutant CpA/K124A/K124Q proteins, using LiCor Odyssey linear quantification software. Bars indicate standard error of the mean ratio. I = Input and U = Unbound. b Cells expressing HA-tagged CpA, K124A or K124Q mutants had their chromatin fibers extracted, and immuno-stained against HA (green) and CENP-C (red) to look for enrichment or depletion of CENP-C on the chromatin fiber (DAPI). Fibers with at least 50% of CENP-C co-localized to the CpA/K124A/K124Q foci were counted and percentage of co-localizing fibers determined. Distribution of the co-localizing fibers is in Additional file 6: Fig. S4C. c ‘Clumping’ K124Q interacts robustly with CENP-C on the chromatin fiber