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. Author manuscript; available in PMC: 2008 Oct 27.
Published in final edited form as: FEBS J. 2007 Nov 26;275(1):138–147. doi: 10.1111/j.1742-4658.2007.06184.x

Fig. 3.

Fig. 3

Submitochondrial localization of RDH13. (A) Mitochondria were fractionated into intermembrane space (IS), outer membranes (OM), matrix (MX) and inner membranes (IM). One-fiftieth of each fraction was separated by SDS-PAGE and the distribution of RDH13 was determined by western blotting. (B) Mitoplasts (mpl) were prepared by hypotonic or digitonin treatment of mitochondria (mch) and incubated with NaCl/Pi, NaCl, Na2CO3 or Triton X-100 (Triton). Treated samples were centrifuged and the distribution of RDH13 between soluble and insoluble fractions was analyzed by western blotting. P, pellet; S, supernatant. The results were identical for digitonin- and hypotonically prepared mitoplasts. (C) Mitochondria or mitoplasts were incubated with the indicated amounts of trypsin (μg) for 30 min on ice, followed by the addition of soybean trypsin inhibitor. RDH13 protein stability was monitored by western blotting.