Fig. 4

Tas2r138 competitively bound to antagonists of PPARG. a To assess potential interaction between AHL and TAS2R138, WT or Tas2r138 siRNA-transfected neutrophils were infected with PAO1 (MOI: 5:1) or AHL-12 (50 µM) stimulated for 2 h. The LDs (stained by Nile red) and PLIN2 (green) expression were detected by western blotting (arrows showing the binding bands). b Dynabeads were incubated with the TAS2R138 antibody overnight at cells 4 °C and decant by the magnetic stand. Neutrophils (1 × 10⁷) were lysed with RIPA and protease inhibitor cocktail and incubated with beads overnight at 4 °C with or without AHL-12-FITC (300 µM). Then the proteins conjugated with beads were isolated by the magnetic stand and measured by western blotting. c The structure of TAS2R138 was modeled by the Swiss model: https://swissmodel.expasy.org/interactive/?ac=Q7TQA6, and the docking analysis was performed on AutoDock platform. d WT or R64A point mutation plasmids were transfected to neutrophils for 24 h. Cell lysate was co-cultured with AHL-12 and TAS2R138 was detected by western blotting (arrows showing the binding bands). e, f WT or Tas2r138 siRNA-transfected neutrophils were infected with PAO1 (MOI: 5:1) or AHL-12 (50 μM) stimulated for 2 h. The PPARG (red) and PLIN2 (green) expression was detected by immunofluorescence imaging (arrows showing typical nuclear distribution and cytoplasmic distribution of PPARG)