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. 2020 Oct 6;11:587913. doi: 10.3389/fimmu.2020.587913

Figure 2.

Figure 2

Lactate attenuates macrophage NF-κB activation and nuclear translocation. Macrophages (Raw 267.4) were treated with lactate 1 h prior to LPS stimulation for 1 h. The cells were harvested for the isolation of the cytosolic and nuclear proteins. (A, B) Lactate (10 mM) suppresses LPS increased levels of NF-κB subunit p65 and its phosphorylation in the cytosol (A) and nuclear translocation (B). (C) Immunostaining shows that lactic acid (10 mM) or sodium lactate (10 mM) suppresses LPS stimulated NF-κB p65 (red) nuclear translocation. Nuclear was stained with DAPI (blue) (Scale bar, 10 µm). (D) Raw 264.7 cells were treated with lactic acid (10 mM) for 1 h and p-IKKα/β and total IKK were assayed by Western blot. (E, F) Lactate suppresses NF-κB activation and nuclear translocation in LPS treated bone marrow derived macrophages (BMDMs). BMDMs were treated with lactate 1 h before the cells were stimulated with LPS. The levels of NF-κB subunit p65 and its phosphorylation in the cytosol (E) and in the nucleus (F) were measured by western blot. N/group = at least three independent experiments. Values are mean ± SD. P values were made by comparison with indicated groups (*P < 0.05; **P < 0.01; ***P < 0.001).