Fig. 6. Downregulation of Rab5a impaired the capacity of the phagosomes to fuse with other organelles. Neutrophils were treated with Rab5a-antisense (AS) or -sense (S) oligonucleotides as described in Materials and Methods. (A) The cells were subsequently lysed and equal amounts of the proteins were separated by SDS-PAGE and immunoblotted with the antibodies raised against Rab5a, actin, hck (azurophil granules), lactoferrin (secondary granules) and LAMP-1 (late endosomes) proteins. (B) Antisense- or sense-treated neutrophils (1x10⁸ cells) were allowed to ingest H37Rv at the ratio of one cell to 20 H37Rv or to 10 S. aureus for 30 minutes. Thereafter, the phagosomes were isolated and equivalent amounts of the phagosomal proteins were separated by SDS-PAGE and immunoblotted for the expression of Rab5a, syntaxin-4, hck, lactoferrin and LAMP-1 using appropriate antibodies. The blots shown are representative of three experiments.