Fig. 5
IFN gene expression and STAT1 phosphorylation, induced by DFX, are mediated by ROS. a, b Quantitative reverse transcriptase PCR analysis for OAS1, OAS3, IFIT1, ISG15, IRF7, EIF2KA, IFITM3, IFITM2 and IFITM1 in Kasumi-1 (a) and K562 (b) cells treated with 100 μM DFX for 24 h. The values are the mean ± SEM of normalized transcript levels of three independent experiments. Per gene evaluated, fold change value of the transcript level in DFX-treated cells compared to untreated cells (CTRL) is reported inside the bar (*p < 0.05 vs CTRL; **p < 0.01 vs CTRL). c, d: Percentage of expression-inhibition of IFN-related gene transcript levels in Kasumi-1 (c) and K562 (d) cells treated with 100 μM DFX + 10 mM NAC, compared to 100 μM DFX only treatment (*p < 0.05 vs DFX-only-treated cells; **p < 0.01 vs DFX-only-treated cells). e, f Western blot analysis of total STAT1 and pSTAT1tyr701 on whole protein Kasumi-1 (e) and K562 (f) cell extracts. Cells were treated with 100 μM DFX ± 10 mM NAC for 24 h. Per cell type, the panel on the top shows representative blots and bar histograms on the bottom represent densitometric analysis showing mean ± SEM of total STAT1 normalized to β-ACTIN (black bars) and p-STAT1/STAT1 ratio (white bars) of three independent experiments (*p < 0.05 vs CTRL; **p < 0.05 vs DFX)