Stable knockdown of CHIP in mpkCCD cells alters AQP2 membrane targeting and ubiquitylation. (A) Representative immunoblots of apically biotinylated AQP2 in control or A4delCHIP cells under control, dDAVP (AVP), or dDAVP followed by 20 minutes washout (AVP washout) conditions. Samples are on the same immunoblot with the same exposure time, and the break is for illustration purposes only. (B) Summary of biotinylation data from five independent experiments. (C) Representative immunoblots of ubiquitylated AQP2 levels in control or A4delCHIP cells under control, dDAVP (AVP), or dDAVP followed by 20 minutes washout (AVP washout) conditions. Samples are on the same immunoblot with the same exposure time, and the break is for illustration purposes only. Arrows represent nonspecific IgG bands. (D) Summary of ubiquitylation data from three independent experiments. Asterisks indicate significance relative to control conditions within an individual cell line; hashtags indicate significance for a specific condition between cell lines. *0.01<P<0.05; **0.001<P<0.01; ***P<0.001; #0.01<P<0.05; ##0.001<P<0.01.