Journal: Cellular and Molecular Life Sciences: CMLS
Article Title: GPR35 prevents drug-induced liver injury via the Gαs-cAMP-PKA axis in macrophages
doi: 10.1007/s00018-025-05751-4
Figure Lengend Snippet: GPR35 attenuates inflammatory response in macrophages through the Gαs-cAMP-PKA pathway. ( A ) Co-IP detection of GPR35 and Gαs in peritoneal macrophages stimulated with or without KA. ( B ) Levels of cAMP in peritoneal macrophages pretreated with FSK, KA, or DSCG. GPR35 agonist KA (4 mM), or DSCG (200 µg/ml) was administered 1 h before treatment with LPS. Adenylate cyclase activator FSK (10 µM) was used to stimulate cAMP in peritoneal macrophages 1 h before treatment with LPS. ( C ) Levels of TNF-α and IL-1β in supernatants from peritoneal macrophages after stimulation with LPS in the presence of FSK, KA, or DSCG. ( D ) Levels of TNF-α and IL-1β in supernatants from peritoneal macrophages with indicated pretreatment after LPS stimulation. ( E ) Levels of TNF-α and IL-1β in supernatants from peritoneal macrophages with indicated pretreatment after LPS stimulation. Gαs inhibitor NF449, AC inhibitor SQ22536, or PKA inhibitor H89 was added to the culture 30 min before treatment with GPR35 agonist KA or DSCG. The data are representative of three independent experiments and shown as the mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001
Article Snippet: To investigate the role of Gαs-cyclic AMP-protein kinase A (Gαs-cAMP-PKA) in the GPR35-mediated effect on macrophages, the Gαs inhibitor NF449 (25 µM, Santa Cruz Biotechnology, TEX, USA), AC inhibitor SQ22536 (50 µM, MedChemExpress, NJ, USA), or PKA inhibitor H89 (10 µM, MedChemExpress, NJ, USA), PKA inhibitor fragment [ – ] amide (10 μm, TargetMol, BOS, USA) was added to the culture 30 min prior to treatment with the GPR35 agonist.
Techniques: Co-Immunoprecipitation Assay
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