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. 1997 Apr 7;137(1):247–258. doi: 10.1083/jcb.137.1.247

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Subcellular colocalization of vacuolar H⁺-ATPase and c-src. Cells maintained on tissue culture plates for 3 d were lysed (S ₀), and the lysate was centrifuged at 1,000 g to remove nuclei and large cell debris. Supernatant was centrifuged at 27,000 g and the membrane pellet (P ₃) was fractionated on a sucrose density gradient. An equal amount of protein from each gradient fraction was subjected to immunoblot using antibodies to c-src, H⁺-ATPase, and F4/80. GLF, Golgi light; GHF, Golgi heavy; CVCF, crude vesicle fractions.