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Effects of DiOHF and Quercetin on Cytochrome c Reduction in DMSO-Differentiated HL-60 Cells

This study examines the impacts of DiOHF and Quercetin on the reduction of cytochrome c in DMSO-differentiated HL-60 cells. The cells were treated with 200 µM cytochrome c and PMA, followed by monitoring the absorbance change (ΔA550-650) over 90 minutes. Compounds were introduced at the 10-minute mark, with H2O2 added as a positive control at 70 minutes. Additionally, Western blot analysis demonstrated the increased expression of p47phox and p67phox proteins following 3 days of DMSO differentiation, while Nox2 levels remained low.

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Effects of DiOHF and Quercetin on Cytochrome c Reduction in DMSO-Differentiated HL-60 Cells

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  1. Supplemental Figure 1 H2O2 (1 mM) 1.4 DMSO control 1.2 DiOHF (30 µM) 1.0 Quer (30 µM) 0.8 A550-650 0.6 0.4 0.2 0 0 10 20 30 40 50 60 70 80 90 min Effects of DiOHF and quercetin on prereduced cytochrome c. DMSO-differentiated HL-60 cells (2 x 107) were washed with Hank’s balanced salt solution (HBSS), resuspended in 1 ml of HBSS containing 200 µM cytochrome c and 100 ng/ml PMA, and incubated for 1 hr at 37oC. Then the cells were removed by centrifugation and the supernatant was transferred to a 96-well plate (200 µl per well). The level of reduced cytochrome c was monitored by the difference of absorbance between 550 nm and 650 nm (A550-650) over 90 min. Compounds were added at 10 min (arrow). At 70 min, 1 mM H2O2 was added to oxidize cytochrome c as a positive control. Data are the mean of triplicate samples.

  2. Supplemental Figure 2 DMSO - + p47phox p67phox Nox2 -tubulin Western blots showing that differentiation of HL-60 with DMSO (1.5%) for 3 days significantly increased the expression of p47phox and p67phox, whereas the level of Nox2 remained low.

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