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. Author manuscript; available in PMC: 2012 Apr 1.

Published in final edited form as: Biochem J. 2011 Apr 1;435(1):175–185. doi: 10.1042/BJ20100840

A). UHRF1 is a multi-domained protein with ubiquitin, PHD, SRA and RING finger domains. The location of UHRF1 siRNAs: si-A, si-B, and si-C target are shown B). Western immunoblot of HCT116 protein lysates from cells transfected with non-targeting siRNA (NTS) or UHRF1 targeting siRNA (si-A, si-B and si-C), show efficient knockdown of UHRF1 with targeting siRNAs. C). Depletion of UHRF1 leads to a G2/M arrest. Percentage of cells in G1 (hatched), S (black) and G2/M (white) in cells transfected with NTS, si-A or si-B. Error bars represent standard deviations from the mean of three experiments. D) Enhanced serine 10 phospho histone H3 levels in UHRF1 knockdown cells. Depletion of UHRF1 (lanes 2 and 3) leads to increased levels of phosphorylated histone H3 (panel 2). Total histone H3 levels are constant. E) There is no concurrent G1 arrest in UHRF1 depleted cells. The remnant G1 population in UHRF1 containing and deficient cells (top histograms) progress to the nocodazole-induced mitotic block (bottom histograms). F) The level of G1 and S cyclins: D2, E and A remain stable in UHRF1 depleted cells (top three panels) while cyclin B1 level is elevated (panel 4). β-actin is used as a loading control for all experiments.

A). UHRF1 is a multi-domained protein with ubiquitin, PHD, SRA and RING finger domains. The location of UHRF1 siRNAs: si-A, si-B, and si-C target are shown B). Western immunoblot of HCT116 protein lysates from cells transfected with non-targeting siRNA (NTS) or UHRF1 targeting siRNA (si-A, si-B and si-C), show efficient knockdown of UHRF1 with targeting siRNAs. C). Depletion of UHRF1 leads to a G2/M arrest. Percentage of cells in G1 (hatched), S (black) and G2/M (white) in cells transfected with NTS, si-A or si-B. Error bars represent standard deviations from the mean of three experiments. D) Enhanced serine 10 phospho histone H3 levels in UHRF1 knockdown cells. Depletion of UHRF1 (lanes 2 and 3) leads to increased levels of phosphorylated histone H3 (panel 2). Total histone H3 levels are constant. E) There is no concurrent G1 arrest in UHRF1 depleted cells. The remnant G1 population in UHRF1 containing and deficient cells (top histograms) progress to the nocodazole-induced mitotic block (bottom histograms). F) The level of G1 and S cyclins: D2, E and A remain stable in UHRF1 depleted cells (top three panels) while cyclin B1 level is elevated (panel 4). β-actin is used as a loading control for all experiments.