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Antibodies Affinity-purified rabbit polyclonal antibodies against the V-ATPase C1,C2, G1, G3, a1, a2, a4, d1, and d2 subunit isoforms were used. These antibodies have been characterized previously. An affinity-purified polyclonal antibody raised in chicken against the V-ATPase E2 subunit was also used to identify narrow and clear cells. A novel affinity-purified rabbit polyclonal antibody against the last 10 amino acids (CMQNAFRSLE) of the C-terminal tail of the V-ATPase A subunit was also used and was characterized in this study. A monoclonal mouse anti-TGN38 antibody was purchased from BD Transduction Laboratories to identify the trans-Golgi network. The following affinity-purified secondary antibodies were used, as appropriate: 1) a goat antirabbit IgG conjugated to fluorescein isothiocyanate (FITC) (Kirkegaard and Perry Laboratories, Gaithersburg, MD), 2) a donkey anti-chicken IgY conjugated to indocarbocyanine (Cy3) (Jackson ImmunoResearch Laboratories, West Grove, PA), 3) and a donkey anti-mouse IgG conjugated to Cy3 (Jackson ImmunoResearch Laboratories).