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Fig. 5 | Plant Methods

Fig. 5

From: Tagging and catching: rapid isolation and efficient labeling of organelles using the covalent Spy-System in planta

Fig. 5

Quantification of isolated chloroplasts and mitochondria bound to SpyCatcher beads after purification. a Quantification of isolated chloroplasts from plants transiently expressing Plastid-SpyTag or wildtype plants. Chloroplasts in direct proximity to beads were counted based on representative microscopic images and the ratio of chloroplasts per bead was calculated. b The enrichment of mitochondria from plants transiently expressing Mito-SpyTag or wildtype plants was determined by marker enzyme assays. The ratio of the specific activity from different marker enzymes of the input fraction (crude plant filtrate before incubation with beads) and the bead fraction (beads after incubation with plant filtrate and washing steps) was calculated. GAP-DH: NADP-dependent glyceraldehyde-3-phosphate dehydrogenase, chloroplast marker; ADH: alcohol deydrogenase, cytosol marker; G6P-DH: NADP-dependent glucose-6-phosphate dehydrogenase, cytosol marker; Fumarase, mitochondria marker)

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Plant Methods

ISSN: 1746-4811