Fig. 1: ChIP-CryoEM technique for the cellular UV-DDB-NCP complex.
From: Structural basis of cyclobutane pyrimidine dimer recognition by UV-DDB in the nucleosome
a U2OS/KO/FLAG-DDB2 cells were immunostained by an anti-DDB2 antibody (green) or an anti-CPD antibody (red). DNA was stained by DAPI (cyan). b Workflow of the ChIP-CryoEM method. c Soluble (Input: lanes 2,3) after MNase treatment, Unbound (Unb.: lanes 4,5), and bound (Eluate: lanes 6,7) fractions after an incubation with anti-FLAG beads were subjected to SDS-PAGE and stained with Oriole (Bio-Rad). Non-treated cells or UV-irradiated cells are indicated as UV- (lanes 2,4,6) or UV+ (lanes 3,5,7). d Amounts of CPDs were measured by ELISA, using an anti-CPD antibody. Non-treated cells and UV-irradiated cells were used as control NCPs and damaged NCPs, respectively. The Y-axis represents the absorbance at 490 nm, normalized to the minimum value across all measurements, which was set to zero. The graph was generated using Prism 10 software, and all data are based on three technical replicates and are presented as mean ± s.d.
