HOXA10-AS does not act as a local regulator of nearby genes and exerts its effects by inducing NF-κB target genes. (A) Expression of HOXA10-AS, HOXA10, HOXA9, and mir-196b as measured by qRT-PCR in EOL-1 cells, MOLM-13, MV4-11, and NB-4 cells after shRNA-mediated HOXA10-AS knockdown using 3 different shRNAs (n ≥ 2; n.d., not detectable). (B) Expression of HOXA10, HOXA9, and mir-196b as measured by qRT-PCR in single-cell clones of MOLM-13, MV4-11, and NB-4 cells after sgRNA-mediated HOXA10-AS excision using 4 different pairs of sgRNAs (n = 2). (C) Enrichment of HOXA10-AS in the cytoplasmic, nuclear soluble, and chromatin-bound compartments as measured by qRT-PCR on fractionated RNA from MOLM-13 and EOL-1 cells. The enrichment is shown as a ratio normalized to the compartment in which the transcript is most highly expressed (n = 2). (D) Gene set enrichment analysis results for NF-κB target genes in LBid-HOXA10-AS–transduced, sorted CD11b⁺/CD14⁻/CD117⁻ cells compared with LBid-control–transduced cells on day 5 of monocytic differentiation. (E) Percentage of phosphorylated p65-positive (pSer529) LBid-HOXA10-AS– or LBid-control–transduced ML-2 cells in the presence or absence of 1 µM BMS-345541 (n = 2; presented as mean ± SD). FDR, false discovery rate; NES, normalized enrichment score.