Figure 6: AtPAM16–GFP localizes to mitochondria inner membrane.
From: Mitochondrial AtPAM16 is required for plant survival and the negative regulation of plant immunity
(a) Immunodetection of AtPAM16–GFP from isolated mitochondria treated with proteinase K. Intact mitochondria (top panel) or sonication-ruptured mitochondria (lower panel) were treated with 10 mg ml−1 proteinase K (PK). Cytochrome c, a known mitochondria inner membrane protein, was used as positive control. (b) Immuno-TEM using a gold-conjugated anti-GFP antibody to detect AtPAM16–GFP. Arrowhead points to mitochondrion, circles highlight gold particles, scale bar represents 200 nm. (c) Quantification of mitochondrial signal relative to background signal in cytoplasm, other organelles and the cell wall revealed significantly more gold per μm2 in mitochondria (ANOVA, P<0.0001, n=81 AtPAM16–GFP, n=94 wild type). * indicates a statistically significant difference, error bars represent s.e. (d) Col-0 seedlings without the AtPAM16–GFP transgene treated with anti-GFP. Arrowheads point to mitochondria, circles highlight gold particles, and scale bar represents 200 nm. (e) The AtPAM16–GFP inner mitochondrial membrane signal was not detected in AtPAM16–GFP seedlings treated without a primary antibody. Arrowhead points to mitochondrion, circles highlight gold particles and scale bar represents 200 nm.
