BMC Genomics 2005, 6:125 http://www.biomedcentral.com/1471-2164/6/125 A. B. 2.5 2 ) (U1.5 H D G 1 0.5 0 C. 50 kDa 50 kDa LAMP-1 γ rTS ß MnSOD 5 10 15 Fractions 20 2.5 2 1.5 1 0 0.5 1.18 n1.14 ) o i t c a r f / g m (1.08 n i e t o r P 1.02 LAMP-1 γ rTS ß MnSOD ) l m / g ( y t i s n e D LFoigcualrizeat3ion of rTSβ and rTSγ to mitochondria Localization of rTSβ and rTSγ to mitochondria. A. Presence of rTSβ in both cytosol and organelle fractions. Five µ g of protein from the organelle (lane 1), cytosol (lane 2), or 5 µ g each of the organelle plus cytosol (lane 3) were resolved by gel electrophoresis, blotted and probed for the presence of the indicated proteins. B. Gradient fractionation of mito- chondria and lysosomes. The crude organellar suspension from CCRF-CEM cells was fractionated on a linear iodixanol gra- dient as described. Individual fractions were collected and assayed for GDH activity and protein content. —, Density (g/ml) of fractions from a mock 5–20% iodixanol gradient; - -, GDH activity; -❍ -, Protein content. C. Western blot. Five µ l of each fraction was denatured and processed for Western blotting as described. Page 6 of 10 (page number not for citation purposes)