Fig. 2

VEN plus MEN1i impaired KMT2Ar-AML growth via repressing HIF-1A-induced HDAC9. A After treating KMT2Ar-AML cell lines THP-1, MV4-11, MOLM13 with DMSO, VEN, MI-503 and their combination for 72 h, RNA sequencing was performed and differentially expressed genes (DEGs) were analyzed; Overall strategy for screening key DEGs related to the synergistic mechanism of VEN plus MI-503 in KMT2Ar-AML was shown; Specific DEGs in VEN plus MI-503 group vs. DMSO group shared by three KMT2Ar-AML cell lines were listed; B The mRNA expression of HDAC9 in THP-1, MV4-11, and MOLM13 after treatments with DMSO, VEN, MI-503, and their combination [two-tailed Student's t-tests; **P < 0.01, ***P < 0.001 ****P < 0.0001]; C The protein expression of HDAC9 and HIF1A in THP-1, MV4-11, and MOLM13 after treatments with DMSO, VEN, MI-503 and their combination; D IC50 of TMP-269 in THP-1, MV4-11, MOLM-13, U937, HL-60, Kasumi-1, and OCI-AML3 cells (72 h); E Growth inhibition was mediated by HDAC9 knockdown in THP-1, MV4-11, and MOLM13 cells; F Different therapeutic effects of VEN plus MI-503 in normal control or HDAC9 knockdown THP-1, MV4-11, and MOLM13 cells were exhibited; Relative absorbance value (optical density [OD] ratio) to normal control for each group was presented and the difference value of OD ratio between two groups was marked as numbers between the histogram bars; G Hypoxia pathway gene set was enriched in the VEN plus MI-503 group compared to DMSO group, the horizontal axis represents the p-value, the vertical axis represents the standard enrichment fraction (NES), the size of the circle represents the -log (p adjusted) value, and different colors represent different gene sets; H The protein expression of HDAC9 and HIF1A in THP-1, MV4-11, and MOLM13 treated with DMOG. I Under HIF-1A inhibitor BAY 87-2243 treatment, HIF-1A mRNA (left) and protein (middle) levels were evaluated. Furthermore, HDAC9 mRNA was detected in DMSO or VEN (400 nM) plus MI-503 (700 nM)-treated THP1 cells with or without BAY 87-2243 3-day pre-treatment (72 h) [two-tailed Student's t-tests; *P < 0.05, **P < 0.01, ***P < 0.001]. J Under normoxia and hypoxia, THP-1 cells were treated by DMSO or VEN plus MI-503, and cell viability and cell apoptosis were evaluated [two-tailed Student's t-tests; **P < 0.01, ***P < 0.001 ****P < 0.0001]