Fig. 5
Upregulation of myeloid-associated CD85k/LILRB4 and CD33 expression during the acquirement of resistance to DOT1L inhibition. ALILRB4 mRNA expression in SEM and SEMPINO_RES cells cultured for 7 days in the absence (-) or presence (+) of 50 µM pinometostat, as determined by RNA-seq. Values indiacte normalized counts with SD derived from four biological replicates for each cell line and condition. Differences in expression were statistically evaluated using unpaired t-tests; **p < 0.005, ***p < 0.0005. B. Differences in chomatin accessibility at the LILRB4 gene locus between SEMPINO_RES and SEM cells as determined by ATAC-sequencing by 2 biological replicates (on top). Red boxes indicate locations within the LILRB4 gene locus of significantly increased of chromatin accessibility in SEMPINO_RES as compared to SEM cells. In addtionm, ChIPseq tracks are presented showing the presence of H3K79Me2, H3K27Ac, H3K4Me3, KMT2A, and AFF1 at the same locus in and indicated cell line models cultured for 7 days in the absence (-) or presence (+) of 50 µM pinometostat. C. Histograms showing the counts of viable cells positive for CD85k/LILRB4 and D. CD33 protein surface expression of indicated cell line models, as determined by flow cytometry (FACS) analysis. Fluorescence Minus One (FMO) controls were used to determine the cut-off point for the positive cell population. E. Quantification of CD85k/LILRB4 and F. CD33 expression represented as the mean ± SD, determined through either one (RS4;11 cells) or two (SEM cells) independent FACS experiments, each involving biological replicates. Differences in expression were statistically evaluated using unpaired t-tests; *p < 0.05, **p < 0.005, ***p < 0.0005