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. 2008 Aug 11;182(3):587–601. doi: 10.1083/jcb.200709177

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© 2008 Noda et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 7. — TM5b knockdown promotes AQP2 trafficking in MDCK/AQP2 cells. (A–D) MDCK/AQP2 cells were transfected with TM5b siRNA (B) or scrambled control siRNA (A), which was indicated by fluorescein-labeled dsRNA (red), and treated without (top) or with (bottom) forskolin. Pretreatment with H89 was also performed using the cells transfected with TM5b siRNA (D) or the control siRNA (C). Cells were labeled for AQP2 (green). Bars, 10 μm. (E and F) Apical cell surface biotinylation assay using MDCK/AQP2 cells transfected with TM5b siRNA or control siRNA, corresponding to A–D. (E) Biotinylated proteins were precipitated with streptavidin-agarose beads and immunoblotted for AQP2. (F) The densitometric quantification normalized to control siRNA-transfected cells without forskolin treatment. Data represent the mean and SE from three independent experiments. *, P < 0.05 versus control siRNA-transfected cells without forskolin treatment; **, P < 0.05 versus control siRNA-transfected cells pretreated with H89 without forskolin stimulation; ***, P < 0.05 versus control siRNA-transfected cells pretreated with H89 with forskolin stimulation.