HJURP-deposited CENP-A recruits constitutive centromere proteins. (A) LacO/TRE U2OS cells were transiently transfected with LacI-HJURP and constructs expressing LAP (GFP localization and purification)-tagged CENP-C, CENP-M, CENP-N, or CENP-T. Cells were preextracted and fixed 72 h after transfection. The presence of CENP-A was assessed using antibodies against endogenous CENP-A. Insets show the arrays at a higher magnification. (B) LacI control images for LAP-CENP-C–T, indicating that recruitment is never observed in the absence of CENP-A. (C) Graph showing the percentage of doubly transfected (GFP and LacI-HJURP) U2OS LacO/TRE cells with endogenous CENP-A present at the array, which also recruited the indicated constitutive centromere proteins (≥30 cells per condition; error bars represent standard deviation). Bars, 5 µm.