Fig. 5

ROS generation is responsible for SIRT1-mediated inflammation in k562 cells. Here, k562 cells were subjected to 5 μM of NAC and 10 μg/mL of LPS for 4 h. a ROS production in k562 cells was reflected by DCF fluorescence intensity. b–d qPCR was performed to determine the mRNA levels of SIRT1, TLR4, and p65 in k562 cells. e WB was performed to determine the protein levels of SIRT1, TLR4, and p65 in k562 cells. f qPCR was performed to determine the mRNA levels of IL-1β, IL-6, and TNF-α in k562 cells. Gene expression in each group was first normalized to the GAPDH gene, and then normalized to the data of the control group. g SIRT1 subcellular localization in LPS-treated k562 cells detected viaIFA. SIRT1 was stained to red through anti-SIRT1 antibody (red), whereas nuclear DNA was counterstained blue through DAPI (blue). The merged image displays the nuclear localization of SIRT1. h Cell fractionation assay showed the location of SIRT1 in nuclear and cytoplasmic fractions. Data are expressed as mean ± standard deviation. *P < 0.05, **P < 0.01 and ^#P < 0.05, ^##P < 0.01 relative to the control and LPS groups, respectively