![Low SLFN11 expression correlates with lower sensitivity to AraC. (A) Correlation analysis between AraC activity and SLFN11 gene methylation in cancer cell lines from the NCI-60 panel. Data were extracted from CellMiner Cross-Database (CDB) (n = 60). (B) Correlation analysis between AraC activity and Log2 of SLFN11 mRNA expression in cancer cell lines from the NCI-60 panel. Data were extracted from CellMiner CDB (n = 59). (A-B) Statistical analysis was performed using simple linear regression, and P values are shown for deviation of the line slope from 0. (C) U937 cells were treated with the vehicle control (dimethyl sulfoxide [DMSO]) or 1 μM AZA for 24 hours. Protein lysates from U937 cells were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), followed by immunoblotting analysis with the indicated antibodies. GAPDH is shown as a loading control. (D) Clonogenic capability of primary leukemic blasts isolated from patients with AML expressing either high levels of SLFN11 (SLFN11-high, n = 3 for 8 ng/ml AraC and n = 4 for 0.08 and 0.8 ng/ml AraC) or low levels of SLFN11 (SLFN11-low, n = 3) treated with either vehicle control (VC; water) or increasing concentrations of AraC, as indicated (left panel). Data are expressed as the percentage of colony formation over VC-treated cells for each patient (Ctrl) and are shown as mean ± standard error of the mean (SEM). Statistical analyses were performed using Mixed-effects analysis followed by Tukey multiple comparisons test and P values are shown for the highest AraC dose for each group. There was a significant interaction (P = .043), that is, the relationship between concentration and percentage of colony formation was significantly different between patients with AML that were SLFN11-low vs SLFN11-high. (D) Relative SLFN11 mRNA expression, normalized to GAPDH, was assessed by qRT-PCR analysis and is shown for the primary leukemic blasts isolated from patients with AML and used in the clonogenic assays (SLFN11-low, n = 3 and SLFN11-high, n = 4; right panel).](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodneoplasia/1/4/10.1016_j.bneo.2024.100037/1/bneo_neo-2024-000191-gr3.jpeg?Expires=1735471010&Signature=GIafVB4hZ2Ux9ZtA66tXsnMiFfUhd~WnFLjm~9umBqshwzVttSOP27dcxE81639kWlWraOKOqMqaujTNtz5TYieQhG2aGuUTNl936it2gcAwKJKlQoIU2KjNJ0fpK0Lq1lIt9Wpon2DU7gZUNRLuiX3KlDUJyez6KwHCcp-Z873h1XuirpbifCBIfKA9AOJo4b4jdkTVI0CdICRV~9ClbnGP4yKlFayOsMeJc2MCx-8GINbkMnSkR9EGOEZwmM6QgxfEFB0LrgUorQTy4aF4JxUTANtvhPApDIkmVLPvPt1LL9uUViv-Qgc1omymd8GRfegKfjZSu~f-d7hcMsRe5w__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Low SLFN11 expression correlates with lower sensitivity to AraC. (A) Correlation analysis between AraC activity and SLFN11 gene methylation in cancer cell lines from the NCI-60 panel. Data were extracted from CellMiner Cross-Database (CDB) (n = 60). (B) Correlation analysis between AraC activity and Log2 of SLFN11 mRNA expression in cancer cell lines from the NCI-60 panel. Data were extracted from CellMiner CDB (n = 59). (A-B) Statistical analysis was performed using simple linear regression, and P values are shown for deviation of the line slope from 0. (C) U937 cells were treated with the vehicle control (dimethyl sulfoxide [DMSO]) or 1 μM AZA for 24 hours. Protein lysates from U937 cells were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), followed by immunoblotting analysis with the indicated antibodies. GAPDH is shown as a loading control. (D) Clonogenic capability of primary leukemic blasts isolated from patients with AML expressing either high levels of SLFN11 (SLFN11-high, n = 3 for 8 ng/ml AraC and n = 4 for 0.08 and 0.8 ng/ml AraC) or low levels of SLFN11 (SLFN11-low, n = 3) treated with either vehicle control (VC; water) or increasing concentrations of AraC, as indicated (left panel). Data are expressed as the percentage of colony formation over VC-treated cells for each patient (Ctrl) and are shown as mean ± standard error of the mean (SEM). Statistical analyses were performed using Mixed-effects analysis followed by Tukey multiple comparisons test and P values are shown for the highest AraC dose for each group. There was a significant interaction (P = .043), that is, the relationship between concentration and percentage of colony formation was significantly different between patients with AML that were SLFN11-low vs SLFN11-high. (D) Relative SLFN11 mRNA expression, normalized to GAPDH, was assessed by qRT-PCR analysis and is shown for the primary leukemic blasts isolated from patients with AML and used in the clonogenic assays (SLFN11-low, n = 3 and SLFN11-high, n = 4; right panel).
