Fig. 3.

Expression of Rab5 induces an enlarged endosomal compartment and a redistribution of P2X4. GFP-tagged rat P2X4 was transiently transfected into HEK-293 cells in the absence or presence of myc-tagged wild-type Rab5, constitutively active Rab5 (Q79L) or inactive Rab5 (S34N). Top panels are confocal images showing the distribution of GFP fluorescence in single cells, and bottom panels show localisation of Rab5-myc proteins using an anti-myc antibody. Cells were grown on 13-mm glass coverslips, fixed and permeabilised and stained with anti-myc primary antibody (1:5,000 dilution), followed by anti-mouse IgG TRITC secondary antibody (1:100 dilution). Images were acquired using a Zeiss LSM510 confocal microscope and a × 63 oil-immersion objective and are representative of three separate transfection experiments. Expression of Rab5 WT or Rab5 Q79L induces an enlarged endosomal compartment that is stained with GFP and anti-myc