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Fig. 2 | Arthritis Research & Therapy

Fig. 2

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. 2

Soluble urate altered subcellular localization of PDZK1 and ABCG2 in HT-29 and Caco-2 cells. Cells treated with 6 mg/dl soluble urate or 10 mM NaOH for 24 h. a Cells fixed with paraformaldehyde and permeabilized with or without Triton X-100 and stained with BXP21 anti-ABCG2 antibody (red). Nuclei stained with DAPI (blue). Scale bar = 50 μm. b Immunofluorescence staining using an antibody against PDZK1 (red). Nuclei stained with DAPI (blue). Scale bar = 50 μm. c Subcellular distribution of PDZK1 and ABCG2 in HT-29 and Caco-2 cells. Cytoplasmic (Cyto, lanes 1 and 2), nuclear (Nu, lanes 3 and 4), and membranous (Memb, lanes 5 and 6) extracts prepared from cells and used for western blot analyses. Data are presented as the mean ± SEM. *P<0.05 and **P<0.01, compared to control cells; n = 3. GAPDH used as a cytoplasmic fraction marker; Lamin A/C used as a nuclear marker; and Na/K ATPase used as a membrane marker. Cytoplasmic fraction normalized to that of GAPDH, whereas membrane fraction normalized to that of Na/K ATPase. ABCG2 ATP-binding cassette transporter, subfamily G, member 2, DAPI 4′,6-diamidino-2-phenylindole, GAPDH glyceraldehyde-3-phosphate dehydrogenase, PDZK1 PDZ domain containing 1, UA uric acid, HUA high concentrations of uric acid (8mg/dl)

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