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. 2020 Nov 10;48(21):12116–12134. doi: 10.1093/nar/gkaa1003

Figure 7.

Figure 7.

UHRF1 enhances LSH chromatin association during S phase of cell cycle. (A) Western blots showing efficient knockdown of UHRF1 by two specific shRNAs in HeLa cells. (B) Cellular fractionation followed by western blot analysis showing UHRF1 knockdown in HeLa cells impaired LSH chromatin association in S phase. (C) Western blots showing efficient knockdown of UHRF1 in NIH3T3 cells by two specific shRNAs. (D) IF analysis showing knockdown of UHRF1 substantially reduced mid-to-late S phase cells with EdU-colocalized LSH foci. Scale bar, 10 μm. (E) Western blots showing efficient knockdown of UHRF1 in HEK293T cells by a UHRF1-specific shRNA. (F) Modified eSPAN assay showing reduced association of LSH with newly replicated repetitive sequences upon knockdown of UHRF1 in HEK293T cells. ** P< 0.005. (G) Working model illustrating how LSH and UHRF1 work together to promote DNA methylation by DNMT1.