Fig. 3

Localization of AQP2-WT in MDCK cells under unstimulated and forskolin stimulated conditions. MDCK cells transduced with LVs containing the AQP2-WT gene for stably expression of AQP2-WT were incubated with 150 μM forskolin prior to immunostaining for AQP2 and tight junctions, ZO-1. The cells were then subjected to CLSM analysis. (a) Schematic representation of the utilized LV expression cassette. The LV transfer vector incorporates a PGK promoter that drives the expression of a single mRNA encoding both AQP2 and puromycin. (b) Unstimulated MDCK cells expressing AQP2-WT. (c) AQP2-WT in MDCK cells after forskolin stimulation. Scale bars = 20 μm. CMV, cytomegalovirus; cPPT, central polypurine tract; LTR, long terminal repeat; PGK, phosphoglycerate kinase 1 promoter; ψ, packaging signal; RRE, Rev-responsive element; U3 (SIN), Self-inactivating deletion in the U3 region; WPRE, woodchuck hepatitis virus post-transcriptional regulatory element