Ar cells (PBMCs) was also determined, as well as the data showed that no substantial cyof 5-demethyl NOB (100 )-treated peripheral blood mononuclear cells (PBMCs) was totoxicity was detected in PBMCs (Figure 1e). These information supported that 5-demethyl also determined, as well as the data showed that no substantial cytotoxicity was detected in NOBPBMCs (Figure 1e). These information supported that various human AML cells; having said that, no significantly decreased the viability of 5-demethyl NOB considerably decreased cytotoxicity in compound-treated normal PBMCs wascytotoxicity in compound-treated the viability of various human AML cells; on the other hand, no detected.normal PBMCs was detected.Int. J. Mol. Sci. 2022, 23,(a)Figure 1. Cont.(b)Int. J. Mol. Sci. 2022, 23,Int. J. Mol. Sci. 2022, 23,4 of4 of(c)(d)(e)Figure 1. 1. The impact of5-demethyl NOB on leukemia cell viability. (a) (a) The chemical structure of Figure The effect of 5-demethyl NOB on leukemia cell viability. The chemical structure of 5-demethylnobiletin (5-hydroxy-6,7,eight,3,4-pentamethoxyflavone).HL-60, THP-1, THP-1, U937, HEL 5-demethylnobiletin (5-hydroxy-6,7,eight,3 ,4 -pentamethoxyflavone). (b) (b) HL-60, U937, HEL and and K562 cells had been treated with car (0.1 DMSO) or 5-demethyl NOB (20048 h. Cell 48 h. K562 cells were treated with car (0.1 DMSO) or 5-demethyl NOB (200 ) for M) for Cell viability was measured by MTT assay. (c) cells have been treated with car, cytarabine (Ara-C, viability was measured by MTT assay. (c) THP-1 THP-1 cells have been treated with vehicle, cytarabine (Ara-C, 20or 5-demethyl NOB (2000 ) for 24M) 48 h.24 and 48 h. was measured by MTT assay. by 20 ) M) or 5-demethyl NOB (2000 and for Cell viability Cell viability was measured MTT assay. (d) THP-1 cells (2.5 had been 5/mL) wereincubated with 5-demethyl NOB (20 and 40 ) (20 (d) THP-1 cells (two.five 105 /mL) 10 seeded and seeded and incubated with 5-demethyl NOB and 4006 h.for 06 h. Thecells was measuredwascounting viable cells using viable cells staining. for M) The number of number of cells by measured by counting trypan blue utilizing trypan blue staining.were treated with 5-demethyl NOB (one hundred )NOB (one hundred M) for 48 h, and cell viability (e) PBMCs (e) PBMCs were treated with 5-demethyl for 48 h, and cell viability was measured was measured by The data represent the mean SD of meanindependent experiments.GDF-5 Protein MedChemExpress experiments.Cathepsin D Protein MedChemExpress by MTT assay.PMID:25818744 MTT assay. The data represent the 3 SD of 3 independent p 0.01 prepresents important differences when compared with the vehicle-treated group. 0.01 represents significant variations when compared with the vehicle-treated group.two.2.2.two. Effects of 5-DemethylNOB on Cell Cycle Progression in in THP-1 Cells Effects of 5-Demethyl NOB on Cell Cycle Progression THP-1 CellsWe next investigated the impact of 5-demethyl NOB on cell cycle progression in THP-1 We next investigated the impact of 5-demethyl NOB on cell cycle progression in cells using flow cytometry analysis. As shown in Figure 2a , the incubation of THP-1 THP-1 cells 5-demethyl NOB (20 andanalysis. As shown in Figure 2a , the incubation of cells with making use of flow cytometry 40 ) for 242 h resulted inside a significant decrease THP-1 cells with 5-demethyl NOB (20 and accumulation in thehSresulted in a substantial within the cell population within the G1 phase and 40 M) for 242 phase in comparison to lower in theDMSO)-treated cells. Treatmentphase5-demethyl NOB didn’t adjust phase comcell population inside the G1 with and accumulation in the S the cell automobile (0.1 pared to.
