Fig. 5: Dual actions of Brr2 in avoiding nonscheduled ciR-loop accumulation.
From: Dual roles of DEAD-box RNA helicase Brr2 in genome stability regulation
a A working model describing the potential role of Brr2 in ciR-loop regulation. b, c S9.6 IF comparing the abundance of R-loops in the control (β-gal KD) and Hel25E KD cells. Two stable cell lines individually overexpressing the wild-type Brr2 (WT) and its truncated mutant deleted of the helicase domain (ΔH1) were used in the assay. Representative fluorescence images from three independent experiments are shown and the relative nuclear S9.6 signals were quantified and calculated from 100 cells. Dashed line indicates border of the nucleus. Scale bars, 10 μm. P value was calculated by two-sided Student’s t-test. d In vitro unwinding assays with synthetic R-loop constructs validating the unwinding activity of Brr2 and its functional domain. R-loops from the WT and ΔH1 treatment groups were run on two different gels. A schematic diagram describing the experimental setup for the assay is also shown to the left of the representative images.
