Supplementary Figure 4: FH binds to H2A.Z.
From: Local generation of fumarate promotes DNA repair through inhibition of histone H3 demethylation
(a) U2OS cells were expressed with a vector for control shRNA or H2A.Z shRNA and reconstituted with expression of WT rH2A.Z or rH2A.Z (NKLLG). Immunoblotting analyses were performed with the indicated antibodies. Data represent one out of 3 experiments. (b) DR-GFP-expressing U2OS cells with depleted H2A.Z and reconstituted expression of the indicated H2A.Z proteins were transfected with a vector that expressed I-SceI. ChIP analyses were performed with an anti-Flag antibody at the indicated time points after I-SceI transfection. Data represent one out of 3 experiments. (c) DR-GFP-expressed U2OS cells with H2A.Z depletion and with or without reconstituted expression of WT rH2A.Z or rH2A.Z (NKLLG) were transfected with a vector with or without expressing I-SceI. PCR analyses for I-SceI-uncut SceGFP were performed 42 h after transfection, as described in Supplementary Fig. 2f. Representative images of PCR products were shown (left panel). The data represent the mean ± SD (n = 3 independent experiments, right panel). Comparable amount of 0.65 Kb PCR products suggested that the efficiency of DSB production by I-SceI in each cell line is similar to each other.
