Fig 1: Validation of DCK from the haploid screen(A) Western blot analysis of DCK protein expression in cell extracts from untreated gene-trap-mutagenised KBM7 cells and from the expanded pool of resistant gene trap-mutagenised KBM7 cells following treatment with gemcitabine (100nM) and entinostat (1µM for 3 days after which it was diluted to 100nM) for 21 days. (B) Western blot analysis of DCK protein expression in cell extracts from wild type and DCK knockout (KO) PANC-1 and SUIT2 cells, following CRISPR/Cas9 targeted inactivation of DCK. (C and D) To assess the effect of DCK inactivation on cell proliferation (C) wild type (WT) and DCK knockout (KO) PANC-1 cells and (D) wild type and DCK KO SUIT2 cells were plated at a density of 4,000 per well in 96-well microtiter plates, allowed to adhere overnight and incubated for 72 hours with vehicle control, or gemcitabine (0.1µM) or entinostat (5 or 10µM) or the combination of gemcitabine plus entinostat. Cell viability was determined by XTT assays. The data are presented as mean inhibition rates from triplicate wells from three independent experiments ±SE. *** P < 0.001.
Fig 2: Insertion sites in the DCK geneSchematic outline of the unique gene-trap insertion sites (red lines) in the DCK gene in cells exposed to gemcitabine and entinostat. Grey boxes represent exons.
Supplier Page from Abcam for Anti-DCK antibody