Extended Data Fig. 9: ALC1 co-operates with PARP activity to permit association of repair proteins with chromatin.
a, DLD1 BRCA2–/– cells were fractionated and the chromatin-bound proteins were immunoblotted. Cells were treated with 5 µM ola for 4 hrs. Data for DLD1 BRCA2–/– cells are from the same sample, from two different western blots and the histone levels for each blot are shown by the ponceau staining. The data for XRCC1 is representative of 5 biologically independent experiments and the data for NTHL1 and APE1 is representative of 3 biologically independent experiments. b, Immunoblot of whole cell lysates of DLD1 BRCA2-/- cells showing total proteins levels upon ALC1 depletion and PARPi treatment. Cells were treated with indicated PARPi for 4 hrs. Data is representative of two biologically independent experiments. c, UWB1.289 cells were fractionated and the chromatin-bound proteins were immunoblotted. Cells were treated with 1 µM tal for 4 hrs. Data for XRCC1 is representative of 4 biologically independent experiments and the data for APE1 is representative of 3 biologically independent experiments. d, Immunoblot showing expression levels of HA-XRCC1. The blot was performed once to access the expression level of the tagged protein. e, Schematic of the IF experiment. f-g, Representative images (f) and quantification (g) of HA-XRCC1 localization to chromatin upon indicated treatments. Scale bar, 50 microns. Data are mean ± s.e.m. from n = three biologically independent experiments, p-value, unpaired Student’s t-test. For each cell line, the median value upon MMS treatment was normalized to its respective untreated control. Source data are provided.
