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Fig. 4 | Arthritis Research & Therapy

Fig. 4

From: Soluble uric acid increases PDZK1 and ABCG2 expression in human intestinal cell lines via the TLR4-NLRP3 inflammasome and PI3K/Akt signaling pathway

Fig. 4

Soluble urate activated the TLR4-NLRP3 inflammasome, and increased expression of PDZK1 and ABCG2 was regulated by the TLR4-NLRP3 inflammasome. Cells preincubated with the caspase-1 inhibitor Ac-YVAD-CMK (20 μM), the TLR1/2 ligand Pam3CSK4 (5 μg/ml), the TLR4 inhibitor TAK-242 (2 μM), or DMSO-Control for 2 h or with P2X7 inhibitor Brilliant Blue G (50 nM) for 6 h. Cells then incubated with 6 mg/dl soluble urate or 10 mM NaOH for 24 h in continued presence or absence of the inhibitor. Indicated cells incubated with 1 μg/ml lipopolysaccharide (LPS) for 6 h. a b ABCG2, PDZK1, pro-caspase-1, active caspase-1 (p10), NLRP3, TLR2, TLR4, MYD88, and P2X7 measured by western blot analysis. Protein expression normalized to that of GAPDH. c Relative mRNA levels of PDZK1 and ABCG2 determined by RT-qPCR. Data presented as mean ± SEM. *P < 0.05 and **P < 0.01, compared to control cells; n = 3. Ac-YVAD-CMK acetyl-YVAD-chloromethylketone, ABCG2 ATP-binding cassette transporter, subfamily G, member 2, ASC apoptosis-associated speck-like protein-containing a CRAD, DMSO dimethylsulfoxide, GAPDH glyceraldehyde-3-phosphate dehydrogenase, LPS lipopolysaccharide, NLRP3 NOD-like receptor superfamily, pyrin domain containing 3, PDZK1 PDZ domain containing 1, TLR Toll-like receptor, UA uric acid

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