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. 2020 May 26;11(5):394. doi: 10.1038/s41419-020-2609-7

Fig. 4. Pyroptosis stress was invalidated once cAMP increased by Forskolin in vitro.

Fig. 4

15 min pre-treatment of Forskolin was exposed to THP-1 cells followed by MSU model for 12 h. a The intracellular cAMP level in THP-1 cells was verified to rise after Forskolin treatment by cAMP assay kit (n = 4). b Forskolin-induced cAMP elevation suppressed the release of IL-1β in THP-1 cells (n = 4). c The THP-1 cells treated with Forskolin presented a decreased rate of pyroptotic cell death in active Caspase-1 and PI double staining by flow cytometry (n = 4). d Western blotting showed that the enhanced cAMP level attenuated the MSU-induced activation of the NLRP3 signaling pathway in THP-1 cells treated with Forskolin. The relative optical density was exhibited in the supplementary materials (n = 4). e Immunofluorescence assay confirmed that enhanced cAMP level attenuated MSU-induced NLRP3 inflammasome activation, as evidenced by decreased expression and colocalization of NLRP3 and ASC in THP-1 cells treated with Forskolin. NLRP3 protein was marked with Alexa Fluor 488 (Green). ASC protein was marked with Alexa Fluor 647 (Red). DAPI (Blue) was used to mark the nucleus. The data were presented as means ± SDs. One-way analysis of variance (ANOVA) with Tukey multiple comparison test was performed. Compared with vehicle group: #P < 0.05, ##P < 0.01, ###P < 0.001. Compared with MSU group: *P < 0.05, **P < 0.01, ***P < 0.001.