Fig. 4. Localization of V-ATPase in the upper vas deferens (UVD) of intact, solvent-injected and colchicine-injected moths. Injections were delivered at Zeitgeber time 8 (Zt8), when moths have low levels of V-ATPase. Colchicine inhibited apical accumulation of V-ATPase in UVDs dissected at Zt12 and Zt16. V-ATPase was detected with an antibody against its B-subunit followed by Texas-Red-conjugated secondary antibody. The images shown are representative of six UVDs processed after each treatment at each time point. Scale bar, 20 µm.