Fig. 7. GPR35 was essential for the activity of the peptide DP7.

a Representative fluorescence images of GPR35 internalization induced by stimulation with DP7 (40 μg/ml) for 30 min in BMDCs. Control indicates the nonstimulated group. b, c GPR35 was inhibited by transfection of JAWSII cells with an shRNA lentivirus, which were then selected with puromycin for 5 days. After stimulation with the peptide DP7 (40 μg/ml) for 30 min, cells were lysed for the detection of transcription factor expression by RT-qPCR (Data are representative of three experiments with three replicates each; Statistical analyses were done using the t-test.) (b) or Erk1/2 phosphorylation by western blotting (c). Control indicates the nonstimulated group. d A simplified schematic of the mechanisms of action of alum/CpG/DP7 in dendritic cells and macrophages. Bars represent means and SEM. *P < 0.05, **P < 0.01, ***P < 0.001.