Figure 4: In situ biochemistry of TREX1-deficient cells.
From: RPA and Rad51 constitute a cell intrinsic mechanism to protect the cytosol from self DNA
(a) Interaction of YFP-TREX1 with endogenous cytosolic nucleic acids in HeLa cells analysed by FLIM-FRET. The fluorescence lifetime of YFP-TREX1 (right), but not of YFP alone (left), decreases significantly in the presence of Sytox Orange (+SO). The lifetime reduction of YFP-TREX1 is abolished in cells pretreated with DNase I (+SO+D). At least ten cells were measured per experiment. Means and s.d. of three independent experiments. ***P<0.001. (b) FCCS of HeLa cells microinjected with dsDNA488_647N. Mean cross-correlation and s.e.m. of three independent experiments measured at t<60 min. (WT, n=2). *P<0.05; **P<0.01; ***P<0.001. (c) Representative FCCS images. ATTO488 (A488, green), ATTO647N (A647N, red). Rhodamine B-dextran (yellow) marks the microinjected cytosol. Scale bar, 10 μm.
