Fig. 7. The effects of in vivo PtdIns4KII inhibition on the endosomal distribution of internalised EGF. Z-series data from time course experiments were reconstructed in three dimensions and the mean diameter of Rh-EGF-labelled punctae measured as described in the Materials and Methods. Punctae were analysed in single cells (300-800 objects per cell) from at least three cells from three independent time course experiments (i.e. 2.7x10³-7.2x10³ objects per time point). (A) Analysis of mean vesicle diameter (± s.e.m.) in 9E10 and 4C5G-transfected cells. The same cells were also scored for scattered punctae (B) and sub-plasma membrane vesicle clusters (C). (D) Analysis of mean vesicle diameter (± s.e.m.) in mock-transfected and PtdIns4KII siRNA-transfected cells. Rh-EGF treated cells were also manually scored for the presence of scattered punctae (E) and the persistence of small vesicles clustered beneath the plasma membrane (F). For B,C and E,F, a minimum of 63 cells per time point were scored from three independent experiments.