
This study used strategies to block AQP2 trafficking at different cellular locations in LLC-PK1 cells, and monitored VP-stimulated phosphorylation of S256 at these sites by immunofluorescence and western blotting with phospho-specific antibodies. It was found that S256 phosphorylation was no longer increased compared to baseline, regardless of AQP2 localization. Taken together, the data indicate that AQP2 S256 phosphorylation can occur at the plasma membrane, in the TGN, or in cytoplasmic vesicles, and that this event is dependent on the expression of PKA in these cells.
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