Fig. 6: Regulation of Aqp14 intracellullar trafficking in X. laevis oocytes.
From: The vertebrate Aqp14 water channel is a neuropeptide-regulated polytransporter
a Amino acid alignment of the C-terminus of Aqp14 from zebrafish (DrAqp14), seabream (SaAqp14) or Atlantic salmon (SsAqp14). Putative phosphorylation sites by PKC (highlighted in blue) or PKA (highlighted in red) are indicated. b Osmotic water permeability (Pf) of oocytes injected with water (control) or DrAqp14, SaAqp14 or SsAqp14 cRNAs, and exposed to the PKC activator PMA (100 nM) or the PKA activator FSK (100 µM), the latter after 1 h incubation with the phosphodiesterase inhibitor IBMX (100 µM), or to the drug vehicle (DMSO, control). c Representative immunoblot of total and plasma membrane (TM and PM, respectively) protein extracts from oocytes treated as in b using the seabream Aqp14 antiserum. d–f Pf of oocytes injected with water or expressing wild-type DrAqp14 or SaAqp14 (WT), or mutant DrAqp14 or SaAqp14 at the putative PKC and PKA phosphorylation residues, and treated with PMA or IBMX/FSK as in b. e–g Representative immunoblot of TM protein extracts from oocytes injected with each construct showing equivalent expression. In b, d and f, data box and whisker plots with the number of biologically independent oocytes indicated above each plot. *P < 0.05; **P < 0.01; ***P < 0.001, statistically different (unpaired Student’s t-test), with respect to control oocytes, with respect to the WT (in parenthesis), or as indicated in brackets. Uncropped blots in c, e and g are shown in Supplementary Figs. 9 and 10.
