To further characterize the decline in RUNX1 activity and other pattern 2 TFs, including the KMT2A::AFF1 TF target MYC that promotes leukemia survival [25], we performed immunoblot analysis (in bulk) and found that AZD1775 treatment decreased RUNX1 protein expression levels in KMT2A-r RS4;11 and SEM cells, and in several non-KMT2A-r ALL cell lines (Fig. 5a, Additional file 1: Fig. S5a). The gene discussed is AFF1; the disease is acute lymphoblastic leukemia.