FIG 3.

Cytoplasmic, soluble nuclear, and chromatin-bound protein levels in HBO1-deleted HeLa cells compared to control cells. HeLa cells were synchronized for 16 h with 50 ng/ml nocodazole and released from nocodazole-induced cell cycle arrest for 2, 4, or 8 h or harvested without release at 0 h, as indicated, and fractionated into chromatin-enriched, soluble nuclear, and soluble cytoplasmic fractions. Lysed proteins were separated by SDS-PAGE, blotted, and subjected to immunodetection of specific proteins, as indicated. (A to C) Representative images of Western blots. (D to F) Densitometry results of MCM6 protein levels normalized to values for wild-type cells at 0 h and relative to loading controls. (G to I) Densitometry results of ORC2 protein levels normalized to values for wild-type cells at 0 h and relative to loading controls. Data are displayed as means ± standards errors of the means for eight experiments (incorporating 2 control cell lines for each of 2 sgRNAs [sgRNA1 and sgRNA2] and 4 HBO1-deleted cell lines for each of the 2 sgRNAs) and were analyzed by Student’s t test. Related data are displayed in Fig. S3 in the supplemental material, including protein levels of other proteins affected by the loss of HBO1.