(A) Levels of the indicated proteins in WCE of HeLa cells transfected with the plasmids expressing shPARP1 or shNT were analyzed using WB.
(B) Nucleotide and predicted amino acid sequences surrounding the intended Cas9 cleavage site (arrow) in WT PARP1 gene as well as the mutated alleles generated by CRISPR-Cas9 are shown. Deleted nucleotides are indicated by capital letters containing strikethroughs, the omitted nucleotides are marked by consecutive dots, and the premature stop codons due to frameshift mutations are indicated by an asterisk. The loss of PARP1 expression in the HeLa-based KO clone was confirmed using WB.
(C and D) Analysis using WB of the indicated proteins in extracts of parental HeLa (WT), the PARP1 KO cells (C), and WT HeLa cells transfected with HA-ELL2 and treated with DMSO or AZD2281 (D).
(E and F) Nuclear extracts (NEs) were prepared from WT HeLa or PARP1 KO cells (E) or from HeLa cells treated with or without AZD2281 (F) and then subjected to immunoprecipitation with the anti-CDK9 antibody or total rabbit IgG. The precipitates were examined using WB for the indicated proteins.
See also Figure S2.