Fig. 3: SMANTIS KO and RUNX1 KO regulate common target genes.

a Scheme of the CRISPR/Cas9 KO strategy for RUNX1 in THP-1 cells by targeting a region next to exon 3 with a single gRNA. b Western blot analysis with antibodies against RUNX1 and topoisomerase I (TOP1) of control (NTC), RUNX1 knockout (KO), and SMANTIS KO (SMTS KO) in THP-1 cells. Heat map using Z-score of the top 50 differential regulated genes as determined by RNA-seq with (KO) and without (NTC) CRISPR/Cas9-mediated KO of SMANTIS (c) and RUNX1 (d). n = 3. e, f Volcano plots of the differential regulated genes from SMANTIS KO and RUNX1 KO compared to NTC. Log2 fold changes between NTC and the KO are shown on the x-axis, the negative logarithmic p-adjusted value (p.adj) is shown on the y-axis. g Venn diagram of the overlapping differentially regulated genes as determined by RNA-seq of SMANTIS KO and RUNX1 KO. h Comparison of the common up- and down-regulated genes between SMANTIS KO and RUNX1 KO. i Comparison of the changed downregulated accessible peaks (down) after RUNX1 CUT&RUN. Common (C; (NTC and SMANTIS KO)) were not changed. j, k Genomic localization of the RUNX1 binding sites in the genome of common and SMANTIS KO (down) specific binding sites. TSS, transcriptional start site; UTR, untranslated region. l Venn diagram of overlapping differentially expressed genes from SMANTIS KO and RUNX1 KO compared to CUT&RUN downregulated peaks in SMANTIS KO. KO, knockout.