FIGURE 3.

Altering AnkG does not impact whole cell ENaC expression but does alter surface channel number. A, representative Western blots showing total ENaC subunit expression with AnkG OE and KD relative to control. B, summary graph of normalized densitometry from Western blots analyzing the expression levels of AnkG and ENaC and the Na⁺/K⁺-ATPase expression. AnkG KD reduced total AnkG expression levels (0.35 ± 0.10 versus 1.00 ± 0.12, n = 5; p < 0.01). AnkG KD did not significantly affect the expression levels of any ENaC subunits or Na⁺/K⁺-ATPase in mCCD cells. AnkG OE also had no effect on ENaC subunits, Na⁺/K⁺-ATPase, or endogenous AnkG. C, summarized results of the effect of FRTs expressing WT or βS518K (locked) ENaC with AnkG. AnkG OE increases βS518K current (1.55 ± 0.11, n = 11) similarly to WT (1.45 ± 0.05, n = 45), demonstrating that AnkG OE altered ENaC N at the membrane surface.