BDH1 Antibody - #BF0318

Fig. 4: Effects of β-OHB treatment on the regulation of M1 macrophage polarization. RAW264.7 cells were first treated with 10 mM β-OHB for 24 h, then cultured under β-OHB-free culture conditions for 24 and 48 h, and stimulated with 100 ng/ml LPS for 24 h. A Immunoblotting analysis of the Kbhb modification in the RAW264.7 cells (n = 3/group). B–D RT-qPCR analysis of IL-6, IL-12, and iNOS mRNA expression levels in the RAW264.7 cells (n = 3/group). The 9-week-old male C57BL/6 mice were orally administered 3 mg/g KE per day for 3d, and their BMCs were obtained and cultured with LCS for 7d to induce BMC differentiation into BMDMs. E, F RT-qPCR analysis of F4/80 and CD11b mRNA expression levels in the BMDMs (n = 3/group). G Immunoblotting analysis of the Kbhb modification in the BMCs treated with LCS at the indicated times (n = 3/group). Partial BMDMs were stimulated with 100 ng/ml LPS for 24 h. H–J RT-qPCR analysis of IL-6, IL-12, and iNOS mRNA expression levels in the BMDMs (n = 3/group). K BDH1 protein expression in mice liver lysates, HepG2 cells, BMDMs, and PMs. L Lack of BDH1 expression leads to β-OHB accumulation in macrophages, which may induce its Kbhb modification. BMDMs were transfected with either Ad-normal control (NC) or Ad-BDH1, then treated with 10 mM β-OHB for 24 h, followed by stimulation with 100 ng/ml LPS for an additional 24 h. M The intracellular β-OHB levels were measured (n = 3/group). N Immunoblotting analysis of the Kbhb modification in the BMDMs (n = 3/group). O–Q RT-qPCR analysis of IL-6, IL-12, and iNOS mRNA expression levels in the BMDMs (n = 3/group). R, S FCM analysis of the proportion of M1 phenotype (iNOS+/F4/80+) mouse BMDMs (n = 3/group). Data are presented as the mean ± standard deviation. *p