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. 2020 Jun 12;39(28):5124–5137. doi: 10.1038/s41388-020-1346-9

Fig. 6. Loss of FACT causes dissociation of the MCM2-7 helicase complex from chromatin during replication stress.

Fig. 6

a–e Immunoblotting of total cell and chromatin extracts from cells transfected with the indicated siRNA. a Representative blots. The levels of chromatin-bound MCM7 (b), MCM4 (c), and PCNA (d). The band intensities were normalized to β-actin. siControl samples are set equal to 1. e Ratio of chromatin-bound MCM4/7 relative to chromatin-bound PCNA. b–e Mean of two biological replicates ±SD. f Immunoblotting of MCF10A cells treated with 2.5 mM HU for the indicated time points. g–i Immunofluorescent staining of MCF10A cells treated with 2.5 mM of HU for 6 h with washed out soluble proteins before fixation. Quantitation of nuclear fluorescence of 20–50 cells per each condition. Only PCNA-positive nuclei were quantitated for MCM7 (h) and PCNA (i) signal.