Hrs and SNX3 Functions in Sorting and Membrane Invagination within Multivesicular Bodies
Figure 7
MEK1 Cleavage by Anthrax Toxin Lethal Factor
(A and B) Experiments (A) and quantification (B) were as in Figure 4G and 4H, except that cells were treated with Hrs siRNAs. Insets in the lower right are a magnification of the regions shown in the boxes.
(C) Hrs or SNX3 was knocked down, or Hrs was knocked down and GFP-SNX3 overexpressed. The trypsin-nicked Protective Antigen (500 ng/ml) and the Lethal Factor (100 ng/ml) of anthrax toxin were then bound to the cell surface on ice [51], and cells were incubated at 37 °C. Lysates (40 μg) were analyzed by western blotting using antibodies against MEK1 N-terminus to detect MEK1 cleavage by Lethal Factor released into the cytosol.
In (B), each condition is the mean of at least three independent experiments; standard errors are indicated. (A) Scale bar indicates 10 μm.