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. 2021 Dec 21;12:771677. doi: 10.3389/fimmu.2021.771677

Figure 5.

Figure 5

Role of protein phosphatase-2A (PP2A) in mediating rhPRG4’s anti-inflammatory effect in monosodium urate monohydrate (MSU) crystals challenged THP-1 human monocytes. THP-1 monocytes were primed with Pam3CSK4 (1μg/mL) for 24h followed by MSU (200μg/mL) crystals ± rhPRG4 (200μg/mL) ± okadaic acid (OKA) (5nM) for 6h and intracellular pro-interleukin-1 beta (pro-IL-1β), secreted mature IL-1β, NLRP3 protein, and caspase-1 activity were quantified. Intracellular pro-IL-1β, NLRP3 and caspase-1 activity were normalized to total isolated protein. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post-hoc test. ns, non-significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. (A) OKA co-treatment increased pro-IL-1β content in rhPRG4-treated THP-1 monocytes. (B) OKA co-treatment increased secreted IL-1β levels in rhPRG4-treated THP-1 monocytes. (C) NLRP3 protein content did not change as a result of rhPRG4 ± OKA treatments. (D) OKA co-treatment increased caspase-1 activity in rhPRG4-treated THP-1 monocytes.