Fig. 3.

A: siRNA against PP1γ increases NFAT5 transcriptional activity, as in Fig. 1B except that PP1γ siRNAs were transfected. PP1γ siRNAs reduce PP1γ protein abundance by ∼69%. *Signficant difference vs. control at 500 mosM (P < 0.05; n = 3). B: PTG-V5 coimmunoprecipitates with PP1γ, and the coimmunoprecipitation is reduced by high NaCl. Medium bathing HEK293 cells stably expressing PTG-V5 was exchanged for medium at 290 or 500 mosM (NaCl added) for 30 min. The cell lysates were immunoprecipitated with the goat anti-PP1γ antibody or goat IgG overnight at 4°C. Immunoprecipitates were probed with mouse anti-V5 and rabbit anti PP1γ. *Signficant difference compared with 290 mosM (P < 0.05; n = 3). C: high NaCl decreases association of PP1γ with PTG-V5 (left) and inhibits activity of remaining PTG-V5-associated PP1γ (right) as in B except that the cell lysates were immunoprecipitated with mouse anti-V5 or IgG. The PP1γ activity was normalized by the amount of PP1γ protein that was co-immunoprecipitated. *Signficant difference compared with 290 mosM (P < 0.05; n = 4). D: high NaCl decreases PTG-V5 abundance. Medium bathing HEK293 cells stably expressing PTG-V5 at 290 mosM was changed to the identical medium or to an otherwise identical one at 500 mosM (NaCl added) for 30 min. Expression of PTG-V5 was determined with mouse anti-V5 antibody. *Signficant difference compared with 290 mosM (P < 0.05; n = 4).