Fig. 3.

Extraction of LDK. (A) Triton X-114 extraction. After incubation with Triton X-114 the samples were separated into aqueous (Aq1) and detergent phase by centrifugation extraction, and the detergent phase was re-extracted (Aq2 and Det). Cell equivalents were analyzed by Western blotting with anti-V5, anti-procyclin, and anti-phosphoglycerate kinase (which detects both the 45-kDa cytosolic and the 56-kDa glycosomal matrix isoforms). (B) Carbonate extraction. Cell equivalents of the carbonate supernatant and pellet in SDS-PAGE sample buffer were loaded onto SDS-PAGE gels and blotted. The sample used for detection of the integral membrane protein VP1 was not boiled but heated to 45°C. Blots were probed with anti-V5, anti-phosphoglycerate kinase (anti-PGK), and anti-VP1. TCL, total cell lysate; P, carbonate pellet; S, carbonate supernatant.