Fig. 8.

Hsc70 influences association of ENaC with endocytic vesicle components. Cells were treated with 0 or 5 μg/ml Dox for 24 h to induce increased Hsc70 expression. Samples underwent immunoprecipitation with anti-Rab7 (A) or anti-Rab11 (B), and β-ENaC was detected in precipitated proteins by immunoblot using anti-V5. Input samples represent 10% of total protein subject to immunoprecipitation. Fold change in V5-β-ENaC coprecipitated with Rab7 or Rab11 was determined by densitometric analysis of precipitated V5 from 4 independent experiments. Values (means ± SE) are presented relative to control (0 μg/ml Dox). *P = 0.029 (A) and 0.018 (B) vs. 0 μg/ml Dox (by t-test). Immunoblots of total Rab7 (A) or Rab11 (B) present in whole cell lysates are included for samples treated with or without Dox and demonstrate that Dox did not alter expression of these species.