Fig. 5.

Soluble urate-induced increases in PDZK1 and ABCG2 expression partially dependent on PI3K/Akt signaling. Cells preincubated with or without the NF-κB inhibitor PDTC (100 μM), the PI3K inhibitor Wortmannin (3 μg/ml), or DMSO-Control for 2 h. Cells then incubated with 6 mg/dl soluble urate or 10 mM NaOH for 24 h. a 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. b d Total protein expression of ABCG2, PDZK1, Akt, p-Akt, and NF-κB p65 determined by western blot analysis. Protein expression normalized to that of GAPDH. c Subcellular distribution of NF-κB p65, PDZK1, and ABCG2 in HT-29 and Caco-2 cells. Cytoplasmic (Cyto, lanes 1, 2, and 3) and nuclear (Nu, lanes 4, 5, and 6) extracts prepared from cells and used for western blot analyses. GAPDH used as a cytoplasmic fraction marker; Lamin A/C used as a nuclear marker. ABCG2 ATP-binding cassette transporter, subfamily G, member 2, DMSO dimethylsulfoxide, GAPDH glyceraldehyde-3-phosphate dehydrogenase, NF-κB nuclear factor-kappa B, PDZK1 PDZ domain containing 1, PDTC pyrrolidinedithiocarbamate, UA uric acid