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. 2001 Mar 20;98(7):3673–3678. doi: 10.1073/pnas.051454398

Figure 3.

Figure 3

Characterization of TGR. (A) Immunoprecipitation. Microsomal fraction was incubated with purified rabbit polyclonal antibodies specific for the C-terminal peptide of TGR in the presence of 100 μM H2O2 (lane 1), water (lane 2), 5 mM DTT (lane 3), 5 mM NADPH (lane 4), or 5 mM GSH (lane 5). TGR was subsequently precipitated with Protein A-Sepharose, and the pellet (precipitated) and supernatant (unprecipitated) fractions were analyzed by immunoblot assays with antibodies specific for TGR. (B) Localization in subcellular fractions. Homogenate of 75Se-labeled mouse testes was fractionated by differential centrifugation. Nuclear (1,000-g pellet), mitochondrial (10,000-g pellet), microsomal (105,000-g pellet), and cytosolic (supernatant after centrifugation at 105,000 g) fractions were analyzed by immunoblot assays with antibodies specific for TGR (Upper) and by PhosphorImager analyses of 75Se on SDS/PAGE gels (Lower). In addition to the TGR location, the location of major 75Se-labeled selenoproteins, TR1, GPx1, and GPx4, are shown on the right. Masses of standards are shown on the left.