Figure 4.
SLFN11 KO AML cells are resistant to AraC but not to HMAs and VEN. (A-B) Protein lysates from U937-Cas9 and U937-SLFN11 KO cells (pooled and single clones, as indicated) were resolved by SDS-PAGE, followed by immunoblotting with the indicated antibodies. GAPDH is shown as a loading control. (C) U937-Cas9 and U937-SLFN11 KO cells (pooled clones) were seeded in 96-well plates and treated with either VC (water) or increasing concentrations of AraC for 24 hours. (D) U937-Cas9 and U937-SLFN11 KO cells (single clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of AZA for 24 hours. (E) Correlation analysis between AZA activity and Log2 of SLFN11 mRNA expression in cancer cell lines from the NCI-60 panel. Data extracted from CellMiner CDB (n = 59). Statistical analysis was performed using simple linear regression, and the P value is shown for the deviation of the line slope from 0. (F) U937-Cas9 and U937-SLFN11 KO cells (pooled clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of DAC for 48 hours. (G) U937-Cas9 and U937-SLFN11 KO cells (single clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of VEN for 24 hours. (H) U937-Cas9 and U937-SLFN11 KO cells (single clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of AZA and VEN for 24 hours. (I) U937-Cas9 control cells and U937-SLFN11 KO cells (single clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of AraC and/or AZA for 24 hours. (C,D,F-I) Cell viability was assessed using the WST-1 reagent. Data are expressed as a percentage of cell viability of VC-treated cells. Means ± SEM of 4 independent experiments in panel C or 3 independent experiments in panels D,F-I are shown. Statistical analyses were performed using a 2-way analysis of variance (ANOVA) followed by the Sidak multiple comparison test. In panel C, there is a significant interaction between cell type and concentration (P < .0001) in AraC-treated cells (the effect of AraC concentration on cell viability was significantly different between U937-Cas9 and U937-SLFN11 KO cells). ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. NS, not significant.

SLFN11 KO AML cells are resistant to AraC but not to HMAs and VEN. (A-B) Protein lysates from U937-Cas9 and U937-SLFN11 KO cells (pooled and single clones, as indicated) were resolved by SDS-PAGE, followed by immunoblotting with the indicated antibodies. GAPDH is shown as a loading control. (C) U937-Cas9 and U937-SLFN11 KO cells (pooled clones) were seeded in 96-well plates and treated with either VC (water) or increasing concentrations of AraC for 24 hours. (D) U937-Cas9 and U937-SLFN11 KO cells (single clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of AZA for 24 hours. (E) Correlation analysis between AZA activity and Log2 of SLFN11 mRNA expression in cancer cell lines from the NCI-60 panel. Data extracted from CellMiner CDB (n = 59). Statistical analysis was performed using simple linear regression, and the P value is shown for the deviation of the line slope from 0. (F) U937-Cas9 and U937-SLFN11 KO cells (pooled clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of DAC for 48 hours. (G) U937-Cas9 and U937-SLFN11 KO cells (single clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of VEN for 24 hours. (H) U937-Cas9 and U937-SLFN11 KO cells (single clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of AZA and VEN for 24 hours. (I) U937-Cas9 control cells and U937-SLFN11 KO cells (single clones) were seeded in 96-well plates and treated with either VC (DMSO) or increasing concentrations of AraC and/or AZA for 24 hours. (C,D,F-I) Cell viability was assessed using the WST-1 reagent. Data are expressed as a percentage of cell viability of VC-treated cells. Means ± SEM of 4 independent experiments in panel C or 3 independent experiments in panels D,F-I are shown. Statistical analyses were performed using a 2-way analysis of variance (ANOVA) followed by the Sidak multiple comparison test. In panel C, there is a significant interaction between cell type and concentration (P < .0001) in AraC-treated cells (the effect of AraC concentration on cell viability was significantly different between U937-Cas9 and U937-SLFN11 KO cells). ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. NS, not significant.

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