Phospho-GRK2 (Ser670) Antibody - #AF3697

Fig. 6 Acetate reduces ERS in neutrophils and inhibits the formation of NETs. a Scanning electron microscopy observation of treated neutrophils (Scale Bar: 2 μm). i: Neutrophil-like differentiation of HL-60 (dHL-60) cells that developed NETs after 4 h of PMA treatment in vitro. ii: There were no NETs in dHL-60 cells treated with acetate and PMA. b Immunofluorescence staining of Sytox-green (green) and MPO (red) in dHL-60 cells, showing that NETs are reduced under sodium acetate treatment (Scale Bar: 50 μm and 25 μm). c Immunofluorescence staining of MPO (green) and p-IRE1 (red) in dHL-60 cells, showing that MPO and p-IRE1 co-localization is reduced under sodium acetate treatment (Scale Bar: 10 μm and 50 μm). d The effects of GRP78/p-IRE1/IRE1/CHOP/ATF6/XBP1/H3cit/MPO in dHL-60 cells exposed to different concentrations of acetate. Data are representative of three independent experiments. e The effects of H3cit/MPO in dHL-60 cells exposed to different concentrations of ethanol. Data are representative of three independent experiments. f The effects of acetate receptor GPR43 and its downstream proteins in dHL-60 cells after stimulated by TG. Data are representative of three independent experiments. g The effects of GRP78/p-IRE1/IRE1/CHOP/ATF6/XBP1/H3cit/MPO in dHL-60 cells after stimulated by TG. Data are representative of three independent experiments. h Immunoprecipitation analysis of ubiquitination of endogenous GRK2 in dHL-60 cells. Data are representative of three independent experiments.

Figure 6 Monomeric derivative of paeoniflorin (MDP) inhibits the expression of hypoxia-inducible factor (HIF)-1α through targeted inhibition of GRK2 S670 phosphorylation. (a) Molecular docking modeling of the compound MDP and G protein-coupled receptor kinase 2 (GRK2), the small molecule, and the critical interaction of 3KRW (human GRK2 in complex with Gbetgamma subunits and balanol) are represented by sticks. (b) A schematic representation of the binding mode of MDP in the GRK2 binding site of 3KRW. (c, d) FLS were treated with or without MDP and exposed to hypoxia for 2 h. Immunoblot analysis and quantification of GRK2, p-GRK2 (S670), and HIF-1α. The values are shown as the mean ± standard error of the mean (n = 3). ∗p < 0.05 compared with the control group and #p < 0.05 compared with the hypoxia group.