Figure 2.

Diosmetin negatively regulates HR repair in EC cells

(a,b): Reporter constructs for DSB repair analysis. HR (a) and NHEJ (b) reporters are constructed and function as previously described [10,26]. (c,d): Analysis of relative HR (c) and NHEJ (d) efficiencies in cells treated by DMSO or diosmetin in indicated concentration. Hec-1B cells were pretreated with DMSO or diosmetin for 24 hours, followed by being transfected with I-SceI digested reporters along with DsRed plasmids. Transfected Hec-1B cells were continued to be treated with the above reagents until 72 hours in the incubator. Cells were collected and examined on the flowcytometry. The ratio of GFP+ versus DsRed+ cells was measured as relative HR or NHEJ efficiency. Students’ t test was used to compare the difference, *: P < 0.05, **: P < 0.01, NS: no significance. Error bars represent SD. (e): Representative pictures of EdU incorporation assay. Hec-1B cells were incubated with DMSO or diosmetin in indicated concentration for 72 hours and subjected to the EdU incorporation assay and then examined on flowcytometry. (f): Statistical analysis of EdU incorporation assay. The distribution of S phase was quantified on the FLOWJO software. At least 10,000 cells were examined. Students’ t test was used to compare the difference. NS: no significance. Error bars represent SD.