G-CSF does not inhibit the release of proapoptotic factors from the mitochondria during apoptosis. Untreated neutrophils or neutrophils treated with 10 ng/mL G-CSF were fractionated at the indicated time points. The cytosolic fractions, which contain no mitochondria, and the pellet fractions, which are enriched for mitochondria, were subjected to SDS-PAGE and analyzed on Western blots stained for the indicated proteins. indicates the full-length proteins and cleavage products in the cases of Bax and Bid. Blots were reprobed for ASC (cytosolic) and MnSOD (mitochondrial matrix) to control for cross contamination and protein loading (bottom panels). All blots are representative of at least 4 independent experiments.