Figure 5. Knockdown of NER and R-loop processing factors reduces E2-induced DNA damage and DSBs.
(A) P-H2AX intensity based on immunostaining of MCF7 cells transfected with indicated siRNA 64 hrprior to the addition of 0 or 100 nM E2 for 24 hr. ****p<0.0001. Western blot shows the level of XPG. (B) Neutral comet assay in cells transfected with indicated siRNA 64 hr prior to the addition of 0 or 100 nM E2 for 24 hr. (C) Quantification of neutral comet tail moment described in (B). ****p<0.0001. >100 comets/condition. (D, E) P-H2AX intensity based on immunostaining in MCF7 cells treated as in (A) with the indicated siRNA. ****p<0.0001. Western blot shows the level of CSB or XPC. The siGL3 control in the quantification shown in (D) is the same as shown in (A). a.u. = arbitrary units. For all data, associated p-values are from non-parametric Mann-Whitney rank sum t-test. n=3 biological replicates. Quantification from >1000 cells/condition unless noted.
Figure 5—figure supplement 1. XPG knockdown does not alter E2-induced cell cycle progression.
