Figure 5.

Detection of Proteins That Cross-React with an At-NEET Antibody in Intact Chloroplasts, Mitochondria, and Plants with Suppressed Expression of At-NEET.

(A) Protein blot analyses of total leaf extract and isolated intact chloroplasts from Arabidopsis (left); total leaf extract, isolated intact chloroplasts, and isolated intact mitochondria from pea (middle); or purified truncated At-NEET or human mitoNEET (right), using antibodies raised against At-NEET.

(B) Detection of organelle-specific proteins in isolated intact chloroplasts and mitochondria from pea performed with antibodies raised against the outer mitochondrial membrane protein VDAC (top panel) or the chloroplast protein PsaD (subunit II of photosystem I; bottom panel).

(C) Protein gel (left) and immunoblot (right) of total leaf extract from wild-type Arabidopsis probed with antibodies raised against At-NEET showing that extraction under denaturing conditions prevents the appearance of the 10-kD reactive polypeptide. LSB, Laemmli sample buffer.

(D) Protein blot showing the expression of At-NEET in the wild type (WT) and the different knockdown and RNAi lines used in this study. RBCS, ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit was visualized with Coomassie blue.