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Fig. 2 | Cell Communication and Signaling

Fig. 2

From: Memantine potentiates cytarabine-induced cell death of acute leukemia correlating with inhibition of Kv1.3 potassium channels, AKT and ERK1/2 signaling

Fig. 2

Memantine potentiates AraC-induced cell death and decline of ATP content and cell proliferation. a Jurkat cells were cultured with AraC±memantine for 72 h and cell death was determined with PI staining and flow cytometry. The mean ± SD percentage of PI+ cells was calculated from n = 4 independent experiments. b Jurkat cells were cultured with AraC and memantine in constant drug ratios for 72 h. The combination index (CI) for AraC+memantine treatment and the dose reduction index (DRI) for AraC was calculated from n = 5 independent experiments with the Chou-Talalay method. c Jurkat cells, cultured in duplicates, were infected with lentivirus harboring shRNA against Kv1.3 channels (Sh-Kv1.3 (1) and Sh-Kv1.3 (2)) or scrambled sequence (Sh-scr). 48 h post lentiviral infection, part of Jurkat cells were treated with 20 nM AraC. SYTOX staining was performed at day 5. The mean + SD percentage of SYTOX+ cells from duplicate cultures is shown for one out of two experiments. d Jurkat cells were cultured without drug, with memantine and AraC±memantine for 72 h and intracellular ATP content was determined with CellTiter-Glo® luminescent assay. Data shows relative light units (RLU) and mean of n = 5 independent experiments. e DNA synthesis of Jurkat cells treated with AraC±memantine was determined by 3[H]-Thymidine incorporation at 72 h; data show mean + SD cpm values of triplicates of one representative experiment (out of 5). f CEM cells were cultured in triplicates with the indicated concentrations of AraC±memantine (100 μM) for 72 h. PI staining was used to determine cell death. The graph shows the mean + SD percentage of PI+ cells. Data is representative for n = 3 independent experiments. g PBMC isolated from a newly diagnosed T-ALL patient (78 years of age, 83% blasts) were cultured in triplicates for 72 h with the indicated concentrations of AraC±memantine. The percentage + SD of sub-G0/1 cells (indicative of dead cells) was determined with PI staining and flow cytometry. h CD3+ T cells were isolated from healthy donors and cultured in triplicates with AraC (3 μM) ± memantine for 72 h. Data show the percentage ± SD of PI+ cells calculated from n = 3 donors. Significance in a-h was determined with Student’s t-test with P* < 0.05, P** < 0.01, P*** < 0.001, and ns = not significant

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