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. 2004 Jan;134(1):182–193. doi: 10.1104/pp.103.031484

Figure 5.

Figure 5.

Western-blot analysis of L. japonicus VDAC expressing yeast and VDACs of L. japonicus and soybean. A and B, L. japonicus VDACs were expressed in a VDAC1-deficient yeast mutant (por-). Approximately 20 μg of total protein was loaded per lane. por+, Strain HR1-2B; por-, strain HR1-2A; 1.1, LjVDAC1.1; 1.2, LjVDAC1.2; 1.3, LjVDAC1.3; 2.1, LjVDAC2.1; 3.1, LjVDAC 3.1. Western blots were incubated with anti-LjVDAC1.1 (A) and anti-POM36 (B) antibody. C to F, Total protein extracts from leaves, roots, and nodules from L. japonicus and soybean and SMs purified via aqueous two-phase partitioning were prepared from 8-week-old plants. Around 20 μg of protein was loaded per lane. L, Leaf; R, root; N, nodule. Western blots were incubated with anti-LjVDAC1.1 (C) and anti-POM36 (E) antibody or with the same antibodies blocked with purified LjVDAC1.1 proteins (D and F, respectively). G, Microsomal fractions of roots and nodules from L. japonicus and SMs from soybean prepared via Percoll density centrifugation were obtained from 8-week-old plants. Around 20 μg of microsomal protein extract and 5 μg of symbiotic membranes were loaded. For comparison, total protein extract of L. japonicus roots and nodules was loaded. VDAC proteins were detected with anti-LjVDAC1.1 antibody. R, Root; N, nodule; t, total protein extract; mf, microsomal fraction. Protein bands representing L. japonicus VDACs are marked with dots. Molecular masses of protein standard are 28 and 39 kD.