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Fig. 6. Association of 120 kDa kinectin isoform with mitochondria in HeLa cells. (A) Western blot of post-nuclear supernatant (lane 1) and mitochondria-enriched fraction (lane 2) of equal concentrations (7 mg/ml), probed with the mitochondrial marker anti-Mn SOD (25 kDa). (B) CT-1 was used to probe the post-nuclear supernatant (lane 1) and mitochondrial fractions (lane 2) of equal concentration (7 mg/ml). 160 kDa and 120 kDa species were detected in lane 1. Only a 120 kDa species was detected in lane 2. (C) The specificity of the affinity-purified anti-insert V2 and V3 antibodies was tested by immunoblotting of a crude cell lysate (concentration 2 mg/ml) Coomassie Blue-stained lysate (lane 1), purified anti-insert V2 and V3 antibodies recognized the prominent 160 kDa form of kinectin (lanes 2 and 3, respectively). (C') Anti-insert V3 antibody was used to probe the post-nuclear supernatant (lane 1) and mitochondrial fractions (lane 2) of equal concentration (7 mg/ml). 160 kDa and 120 kDa kinectin were detected in lane 1 whereas a 120 kDa band was detected in lane 2. (D) An ER marker, anti-calreticulin (63 kDa), was used to probe the post-nuclear supernatant (lane 1) and mitochondrial fractions (lane 2) of equal concentration (0.76 mg/ml). ER protein was not enriched in lane 2.

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