Fig. 2: And-1 plays a critical role in recruiting Polδ-p125 to UV lesion sites to facilitate repair synthesis.

A Mass spectrometry analysis to identify And-1-associated proteins involved in the NER. B, C Co-immunoprecipitation (co-IP) to detect the interaction of endogenous And-1 (B) or exogenous FLAG-And-1 (C) with indicated proteins in HEK293T cells (n = 3 independent experiments). D Chromatin fractions and whole-cell proteins were extracted from HaCaT cells at indicated time points following UVB exposure at 40 mJ/cm² and cell lysates were analyzed by immunoblotting for the indicated proteins (n = 3 independent experiments). E HaCaT cells were transfected with indicated siRNA and plasmid for 40 h, and cells were then exposed to UVB at 75 mJ/cm² and harvested 1 h post UVB irradiation. Chromatin fractions and whole-cell proteins were extracted and immunoblotted for indicated proteins (n = 3 independent experiments). F Immunofluorescence staining was performed to examine the co-localization of p125 and CPDs in HaCaT cells exposed to UVB at 75 mJ/cm². G HEK293T cells were transfected with indicated siRNA and plasmid for 40 h, then harvested 1 h post UVB irradiation at 100 mJ/cm². V5-IP was performed in cell lysates, and then IPs were immunoblotted for indicated proteins. H HaCaT cells were transfected with the indicated siRNA and plasmid for 40 h, then exposed to UVB at 75 mJ/cm² and allowed to repair for 2 h. Unscheduled DNA synthesis (UDS) was then performed to assess Edu incorporation efficiency. DNA damage was identified by CPD staining. EdU levels were quantified by counting approximately 100 cells and normalized to control cells, n = 3 independent experiments. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired two-tailed t-tests in GraphPad Prism 9.0. ***P ≤ 0.001. Source data are provided as a Source Data file.