Cell viability was assessed using the MTS assay (G1111; Promega) following treatment with different concentrations of DAC for 7 days; the optical density was measured at 490 nm. Cell viability was calculated using the following formula: cell viability (%) = (Abstreated blank/Abscontrol blank) × 100%.
After successfully transfecting cells with shNC or shLIN7A lentivirus, the cells were treated with 10 nM DAC for 3 days; the medium was then replaced with fresh medium. On day 4, 2 × 105 cells/well were seeded into 6-well plates and either left untreated or treated with 100 nM Ara-C for an additional 3 days. Thereafter, on day 7, the cells were harvested and stained with annexin V-FITC (BD Biosciences, Franklin Lakes, NJ, USA) for 15 min at room temperature in the dark. Propidium iodide was added to the samples before analysis to distinguish live cells from dead ones, and the apoptosis rate was determined using a CytExpert flow cytometer (Beckman Coulter Life Sciences, Indianapolis, IN, USA). Apoptosis experiments were performed in triplicate using the following treatment groups: DAC alone, Ara-C alone, and DAC followed by Ara-C.
After successfully transfecting cells with shNC or shLIN7A lentivirus, the cells were treated with 10 nM DAC for 3 days; the medium was then replaced with fresh medium. On day 4, 2 × 105 cells/well were seeded into 6-well plates and either left untreated or treated with 100 nM Ara-C for an additional 3 days. Thereafter, on day 7, the cells were harvested and stained with annexin V-FITC (BD Biosciences, Franklin Lakes, NJ, USA) for 15 min at room temperature in the dark. Propidium iodide was added to the samples before analysis to distinguish live cells from dead ones, and the apoptosis rate was determined using a CytExpert flow cytometer (Beckman Coulter Life Sciences, Indianapolis, IN, USA). Apoptosis experiments were performed in triplicate using the following treatment groups: DAC alone, Ara-C alone, and DAC followed by Ara-C.
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