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Additional file 12 : Figure S7. HOTAIRM1 expression in the different transfected cells. a-b HOTAIRM1 expression in HOTAIRM1-silenced (a) and HOTAIRM1-enforced OCI-AML3 cells (b). c-d HOTAIRM1 expression in HOTAIRM1-silenced (c) as well as HOTAIRM1-enforced OCI-AML2 + NPM1-mA cells (d). e HOTAIRM1 expression in HOTAIRM1-enforced OCI-AML2 cells.

Additional file 10 : Figure S5. Enrichment of KLF5 on the E2 fragment of the HOTAIRM1 promoter was measured by ChIP assays in leukemia cells.

Additional file 17 : Figure S12. Screening for the ubiquitin ligase of EGR1. a Network view of the predicted E3 ubiquitin ligase for EGR1. b Recognition motif for the potential E3 ubiquitin ligase MDM2 in the EGR1 sequence.

Additional file 4 : Table S4. The top 10 predicted factors bound to HOTAIRM1 promoter by JASPAR database analysis.

Additional file 16 : Figure S11. EGR1 does not affect the expression of HOTAIRM1. a The efficiency of EGR1 overexpression in OCI-AML3 cells was validated by western blotting. b qRT-PCR was used to detect the expression of HOTAIRM1 in EGR1-enforced OCI-AML3 cells. The data are presented as the mean ± SD of three independent experiments. n.s. indicates no significant difference.

Additional file 3 : Table S3. Primers used for ChIP-qPCR.

Additional file 11 : Figure S6. Co-IP assays were used to evaluate the binding of NPM1 and KLF5, NPM1-wt and WWP1 in OCI-AML3 cells. a-b The interaction between NPM1-mA (a) or NPM1-wt (b) and KLF5 in OCI-AML3 cells were determined by IP assays. c IP assays were used to determine the binding of WWP1 to NPM1-wt in OCI-AML3 cells.

Additional file 19 : Figure S14. Cytoplasmic HOTAIRM1 promotes leukemia cell autophagy and proliferation through ULK3. a, c Western blot analysis of LC3-II, p62 and ULK3 levels in HOTAIRM1-silenced OCI-AML3 cells following ULK3 overexpression (a) and HOTAIRM1-overexpressed OCI-AML2 cells following ULK3 knockdown (c). b, d CCK-8 assays were used to analyze cell proliferation in transfected OCI-AML3 (b) and OCI-AML2 cells (d). The data are presented as the mean ± SD of three independent experiments. *P

Additional file 1 : Table S1. Clinical characteristics of newly diagnosed AML patients.

Additional file 6 : Figure S1. Stable NPM1-mA-GFP expressing OCI-AML2 cells (OCI-AML2 + NPM1-mA) were generated. a Cytoplasmic localization of NPM1-mA in the OCI-AML2 stably expressing Flag-GFP tagged NPM1-mA. Cytospun cells were fixed and stained with DAPI. b The level of NPM1-mA in leukemia cells were measured by western blot analysis.

Additional file 13 : Figure S8. HOTAIRM1 promotes autophagy and proliferation in transfected OCI-AML2 + NPM1-mA leukemia cells. a, i Western blot analysis of LC3-II and p62 levels in OCI-AML2 + NPM1-mA cells after transfection of shHOTAIRM1 (a) and HOTAIRM1 expression vector (i). b, j Representative microscopy images of LC3 puncta in HOTAIRM1-silenced OCI-AML2 + NPM1-mA (b) and HOTAIRM1-enforced OCI-AML2 + NPM1-mA (j). The bar graphs showed the quantification of the fluorescent puncta data. Scale bar: 25 μm. c, k Evaluation of cell viability in OCI-AML2 + NPM1-mA cells transfected with shHOTAIRM1 (c) or HOTAIRM1 plasmid (k) for indicated by CCK-8 assays. d, l Evaluation of cell proliferation in HOTAIRM1-silenced (d) and HOTAIRM1-enforced OCI-AML + NPM1-mA cells (l) for indicated by EdU assays. The bar graphs showed the percentage of EdU positive cells. Scale bar: 100 μm. e, m Flow cytometry was performed to assess cell cycle of HOTAIRM1-silenced (e) and HOTAIRM1-enforced OCI-AML2 + NPM1-mA cells (m). The bar graph shows the percentages of G0/G1-, S-, and G2/M-phase cells. f, n Flow cytometry was used to detect apoptosis of HOTAIRM1-silenced (f) and HOTAIRM1-enforced OCI-AML2 + NPM1-mA cells (n). LL, dead cells; UL, viable cells; LR, early apoptotic cells; UR, late apoptotic cells. g, o Flow cytometric analysis was performed to analyze apoptosis of OCI-AML2 + NPM1-mA cells after transfection with shHOTAIRM1 (g) or HOTAIRM1 plasmid (o), followed by treatment with 200 nM Ara-C for 48 h. h, p The protein levels of Cyclin D1, CDK4, Bax and Bcl-2 in HOweTAIRM1-silenced (h) and HOTAIRM1-enforced OCI-AML2 + NPM1-mA cells (p). The data are presented as the mean ± SD of three independent experiments. **P

Additional file 5 : Table S5. The top 10 proteins with specific binding to HOTAIRM1 identified by mass spectrometry analysis.

Additional file 18 : Figure S13. Nuclear HOTAIRM1 promotes cell cycle progression and inhibits apoptosis in leukemia cells through EGR1. a, c Flow cytometry was used to determine and compare the differences in cell cycle progression in differently transfected groups of OCI-AML3 (a) and OCI-AML2 cells (c). b, d Flow cytometry was used to determine and compare differences in apoptosis in differently transfected groups of OCI-AML3 (b) and OCI-AML2 cells (d). The data are presented as the mean ± SD of three independent experiments. *P

Additional file 7 : Figure S2. The expression levels of HOTAIRM1 in the nucleus and cytoplasm of leukemia cells were measured by qRT-PCR.

Additional file 20 : Figure S15. Cytoplasmic HOTAIRM1 promotes cell cycle progression and inhibits apoptosis in leukemia cells through ULK3. a-b Flow cytometry was used to analyze the changes in cell the cycle (a) and apoptosis (b) in HOTAIRM1-silenced OCI-AML3 cells following ULK3 upregulation. c-d Flow cytometry was used to analyze the change in the cell cycle (c) and apoptosis (d) in HOTAIRM1-overexpressed OCI-AML2 cells following ULK3 downregulation. The data are presented as the mean ± SD of three independent experiments. *P

Additional file 8 : Figure S3. NPM1-mA expression in leukemia cells in the different treatment groups. a The efficiency of RNA interference-mediated knockdown of NPM1-mA expression was determined by qRT-PCR. b The efficiency of RNA interference-mediated knockdown of NPM1-mA expression was determined by western blotting. c NPM1-mA protein levels in NPM1-mA enforced OCI-AML2 cells. d Western blot analysis of NPM1-mA in OCI-AML3 cells treated with KPT-330. β-tubulin as the cytoplasmic control and PCNA as the nuclear control. The data are presented as the mean ± SD of three independent experiments.

Additional file 14 : Figure S9. HOTAIRM1 promotes autophagy and proliferation in transfected OCI-AML2 leukemia cells. a Western blot analysis of LC3-II and p62 levels in OCI-AML2 cells after transfection of HOTAIRM1 expression vector. b Representative microscopy images of LC3 puncta in HOTAIRM1-enforced OCI-AML2 cells. The bar graphs showed the quantification of the fluorescent puncta data. Scale bar: 25 μm. c Evaluation of cell viability in OCI-AML2 cells transfected with HOTAIRM1 plasmid for indicated by CCK-8 assays. d Evaluation of cell proliferation in HOTAIRM1-enforced OCI-AML2 cells for indicated by EdU assays. The bar graphs showed the percentage of EdU positive cells. Scale bar: 100 μm. e Flow cytometry was performed to assess cell cycle of HOTAIRM1-enforced OCI-AML2 cells. The bar graph shows the percentages of G0/G1-, S-, and G2/M-phase cells. f Flow cytometry was used to detect apoptosis of HOTAIRM1-enforced OCI-AML2 cells. LL, dead cells; UL, viable cells; LR, early apoptotic cells; UR, late apoptotic cells. g The protein levels of Cyclin D1, CDK4, Bax and Bcl-2 in HOTAIRM1-enforced OCI-AML2 cells. The data are presented as the mean ± SD of three independent experiments. **P

Additional file 2 : Table S2. Primers used for qRT-PCR.

Additional file 15 : Figure S10. HOTAIRM1 expression in HOTAIRM1-silenced OCI-AML3 cells transfected with different vectors.