Fig. 4

Transcription redistributes MCM in the G1 phase. a Western blotting showing detection of the indicated proteins in G1-arrested K562 cells in the absence or presence of 10 μg/mL α-amanitin. G1-arrested K562 cells were treated with or without 10 μg/mL α-amanitin for 12 h before harvest. WCE, whole cell extract; Cyto-E, cytoplasmic extract; Chro-E, chromatin extract. b The distribution of ORC2 within the ERIZ-associated non-transcribed regions (non-TRs) in G1-arrested K562 cells in the absence or presence of 10 μg/mL α-amanitin. The non-TRs are the same as those shown in Fig. 3a. c Box-plot showing the log2 ratio of the read density of ORC2 between non-TRs and flanked TRs in the A compartments of K562 cells treated with 10 μg/mL α-amanitin. The Wilcoxon rank-sum test was applied for statistical analysis (p = 0.31). d The distribution of MCM5 in G1-arrested K562 cells in the absence or presence of 10 μg/mL α-amanitin. The data from ChIP-ed samples over the input samples are defined as fold change data, as in the ENCODE project and a previous report [35]. The fold change data were normalized to z-scores for heatmaps. The displayed regions are the same as in b. e Box-plot showing the log2 ratio of MCM5 signal enrichment in non-TRs and TRs in the A compartments of K562 cells treated with 10 μg/mL α-amanitin. The Wilcoxon rank-sum test was applied for statistical analysis. f, g, and h The distribution of early replication initiation from EdU/HU (f), ORC2 (g), and MCM5 (h) in ERIZ-adjacent active gene bodies with or without 10 μg/mL α-amanitin treatment. TSS, transcription start site; TTS, transcription termination site. ERIZ-flanked transcribed regions larger than 50 kb are ranked by width (with the smallest genes on top). For display, all transcribed regions were scaled to the same width and aligned at both TSS and TTS. Each line represents an individual transcribed gene