Fig. 4: Genetic perturbation of BRPF1 sensitizes Taxol-resistant T1-160 cells.

a A schematic representation of dual-color competition assay. mCherry-H2B- or eGFP-H2B-labeled T1-160 cells were infected with NT1 or sgRNA of interest. Cells were mixed in a 1:1 ratio, treated with Taxol (160 nM) and imaged after attachment and every 4 days until day 24. The ratio of eGFP-H2B-T1-160 cells carrying the sgRNA of interest was determined to identify the genes whose loss revert resistance. Created with BioRender.com. b Representative images of dual-color competition assay for KOs of indicated genes. Scale bar, 200 µm. c Statistical analysis of eGFP⁺ cells upon ABCB1 and BRPF1 KO after dual-color competition assay. d Complexes that are formed by BRPF and JADE proteins. Created with BioRender.com. e Western blot analysis of BRPF1 protein levels in parental and resistant cells. f Western blot analysis of BRPF1 expression upon BRPF1 KO in T1-160 cells. g Clonogenic assay showing the effect of BRPF1, BRPF2 and BRPF3 KOs in the presence of Taxol in T1-160 cells. h Quantification of the colonies in g. i The effect of BRPF1 siRNA on colony-forming abilities of T1-160 cells in the presence of Taxol. j Quantification of colonies in i. k Cell viability measurement performed with siNT and siBRPF1 samples in the presence of Taxol in T1-160 cells. l The effect of MOZ–MORF complex members’ KOs on colony-forming abilities of T1-160 cells in the presence of Taxol. m Quantification of colonies in l. n BRPF1 protein levels upon KO and overexpression of PAM mutant version of GFP fused BRPF1 in T1-160 cells o, Rescue experiment with overexpression of BRPF1 PAM mutant version on BRPF1-KO cells in the presence of Taxol. p Quantification of colonies in o. P values determined by two-tailed Student’s t-test. For each condition (DMSO or Taxol), statistical comparisons were made relative to their respective control groups. ; *P < 0.05, **P < 0.01, ***P < 0.001.