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. 2022 Jan 11;12(1):5. doi: 10.1038/s41408-021-00603-3

Fig. 1. Depletion or degradation of Menin increases sensitivity to treatment with venetoclax or abemaciclib in AML cells.

Fig. 1

A MOLM13 was transfected with sgRNA against Exon 2 or Exon 6 of Menin or a negative control sgRNA (sgNeg) and incubated for 5 days. Then, total cell lysates were prepared and immunoblot analyses were conducted. The expression levels of GAPDH served as the loading control. B MOLM13 cells were transfected with sgNeg or sg Menin Ex 2 or Ex 6 and incubated for 72 h. Then, cells were treated with the indicated concentrations of venetoclax for 48 h. The % of annexin V-positive, apoptotic cells were determined by flow cytometry. Mean of two independent experiments performed in duplicate ±S.D. **p < 0.01 compared to sgNeg-transfected MOLM13 cells (determined by a two-tailed, unpaired t test). C MOLM13 cells were transfected with sgNeg or sg Menin Ex 2 or Ex 6 and incubated for 72 h. Following this, cells were treated with the indicated concentrations of abemaciclib for 96 h. The % of TO-PRO-3 iodide-positive, non-viable cells were determined by flow cytometry. Mean of two independent experiments performed in duplicate ±S.D. **p < 0.01 compared to sgNeg-transfected MOLM13 cells (determined by a two-tailed, unpaired t test). D MOLM13-Menin-FKBP12F36V cells were treated with the indicated concentrations of dTAG-13 for 72 h. At the end of treatment, cell lysates were prepared and immunoblot analyses were conducted for HA-tagged Menin, endogenous Menin, MEIS1, FLT3, CDK6, PBX3, BCL2, and p27. The expression levels of GAPDH served as the loading control. E MOLM13-Menin-FKBP12F36V cells were treated with the indicated concentrations of venetoclax for 48 h alone or co-treated with 500 nM of dTAG-13. The % of TO-PRO-3 iodide-positive, non-viable cells were determined by flow cytometry. Mean of two independent experiments performed in duplicate ±S.D. **p < 0.01 compared to MOLM13-Menin-FKBP12F36V cells not treated with dTAG-13 (determined by a two-tailed, unpaired t test in GraphPad V8). F MOLM13-Menin-FKBP12F36V cells were treated with the indicated concentrations of abemaciclib for 96 h without or with 500 nM of dTAG-13. The % of TO-PRO-3 iodide-positive cells was determined by flow cytometry. Mean of two independent experiments performed in duplicate ±S.D. **p < 0.01 compared to MOLM13-Menin-FKBP12F36V cells not treated with dTAG-13 (determined by a two-tailed, unpaired t test in GraphPad V8).