Fig. 4. (A) Effects of PEX2 depletion on the location of HXK and PGI activities. Bloodstream and procyclic pex2 cells were disrupted using glass beads. Enzymatic activities were measured in the cytosolic (black) and organellar pellet (gray) fractions. The +Tet columns are from cells grown in the presence of 100 ng/ml Tet for 36 hours; -Tet columns are the controls. Measurements were made with three independent cell extracts; the activities shown are the mean and standard deviation from two to five readings. (B) Western blots showing the distribution of proteins in procyclic mutants, 0, 1, 2 and 3 days after Tet addition. Cells were separated into digitonin-soluble (s) and pellet (p) fractions. The proteins detected are indicated on the right. (C) Western blot showing the expression of EP1 in procyclic mutants grown in the absence and in the presence of Tet for up to 3 days. Each lane was loaded with a total protein extract from 5x10⁵ cells.