(A) Super-resolution imaging was performed in U-2OS cells to analyze the colocalization between XRCC5 (magenta) and FBXL10, RNF68 and RNF2 (green), with or without 20 min of NCS treatment. …
(A) HEK293T cells were transfected with FLAG-RNF68, HA-RNF2, and Myc-FBXL10. Cell lysates were immunoprecipitated with an anti-FLAG resin, followed by elution using 3x FLAG peptide. The eluate was …
(A) Representative images of the kinetic plots showed in Figure 1C. U-2OS cells stably expressing either GFP-FBXL10, mCherry-RNF68 and GFP-RNF2 and transfected with the indicated siRNAs, were …
(A) U-2OS cells stably expressing either GFP-FBXL10 were pre-sensitized with BrdU (10 μM) for 36 hr and subjected to 405 nm laser induced damage. DNA damage recruitment dynamics were captured by …
(A) U-2OS cells stably expressing GFP-FBXL10 (left), mCherry-RNF68 (middle) or GFP-RNF2 (right) were transfected with siRNAs targeting RNF51, RNF110 or a non-targeting control (CTRL). Cells were …
(A) Representative images of the kinetic plots showed in Figure 2A. U-2OS cells stably expressing either GFP-FBXL10, mCherry-RNF68 and GFP-RNF2 and transfected with the indicated siRNAs, were …
Left, Schematic representation of RNF68 mutants used in mapping experiments. RNF68 mutants found to interact with FBXL10 or RNF2 are indicated by the symbol (+). Right, HEK293T cells were …
(A) U-2OS cells were transfected with the indicated siRNAs. Cells were fixed 30 min after laser microirradiation and stained with antibodies to γH2A.X (orange) and H2AK119Ub1 (green). Scale bar …
(A) U-2OS cells were transfected with the indicated siRNA sequences. The graph shows representative knockdown efficiencies of the indicated genes. Corresponding mRNA levels were analyzed using …
(A) Schematic of the U-2OS reporter system that allows monitoring transcription following induction of DSBs. DSBs are induced in the LacO repeats by activating the ER-mCherry-LacI-FokI-DD fusion …
(A) U-2OS FokI-YFP-MS2 reporter cells used to detect transcriptional repression upon DNA damage were transfected with either the indicated siRNAs or a non-targeting control (siCTRL). 48 hr later, …
(A) U-2OS cells were transfected with siRNA oligos targeting FBXL10, RNF68, RNF2, RNF8 or a non-targeting control (siCTRL). Cells were treated for 3 hr (BRCA1, Rad51) or 1 hr (FK2, pRPA S33) with …
(A) U-2OS cells were transfected with siRNAs targeting FBXL10, RNF68, RNF2, RNF8, or a non-targeting control (siCTRL). Cells were treated with 0.1 μg/ml NCS (+) for 1 hr (FK2, pRPA S33, MDC1, …
(A) U-2OS cells were transfected with siRNAs targeting FBXL10, RNF68, or RNF2, or a non-targeting control (siCTRL). Cells were treated with 0.1 μg/mL NCS for 30 min or 1 μM CPT for 3 hr. Cells were …
(A) qChIP was performed in the DSB reporter U-2OS cells (see Figure 3C) with and without FokI-induced DSBs using the indicated antibodies. The results are shown as a ratio of H2A.Z to H2A levels. …
(A) U-2OS FokI reporter cells were used in qChIP experiments. The ER-mCherry-LacI-FokI-DD nuclease was induced for 1 hr using Shield-1 and 4-OHT. qChIP was performed with and without …
(A) U-2OS cells were transfected with H2A.Z siRNA or CTRL siRNA. Cells were fixed 30 min after laser micro-irradiation and stained with antibodies to γH2A.X (orange) and H2AK119Ub1 (green). Scale …
U-2OS FokI-YFP-MS2 reporter cells used to measure transcription near DSB sites were transfected with siRNA against H2A.Z or a non-targeting control (siCTRL).
48 hr later cells were treated with Shield-1 ligand (250 nM) and 4-OHT (500 nM) for 4 hr. Cells were lysed, and immunoblotting was performed as indicated.
After 72 h, cells were treated with NCS for 20 minutes. After harvesting, cells were fractionated into soluble and chromatin fractions, and lysates were immunoblotted as indicated. For chromatin …
Cells were pre-sensitized with BrdU (10 μM) for 36 hours and subjected to 405 nm laser induced damage. DNA damage recruitment dynamics were captured by live cell imaging. Relative fluorescence …
Raw data for all graphs in main figures and figure supplements.