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Figure 3. Metabolic labeling of mutant DProPHMs proteins. Duplicate wells of each cell line were incubated with [ 35 S]Met/Cys for 15 min; one well was extracted immediately (P15) while the other was rinsed and incubated in CSFM containing unlabeled Met/Cys for 120 min before harvesting the medium (Mdm) and extracting the cells (C120). PHM was immunoprecipitated from aliquots representing equal fractions of cell extract and medium, fractionated by SDS-PAGE, and visualized by fluorography. Each cell line was analyzed at least 2 times; for each mutant, two independent cell lines were analyzed.

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