VSV-G transport is impaired in primary cells from mutant PrP but not PrP knockout mice.A, MEFs from WT, CJD, FFI, and PrP knockout (KO) mice were infected with VSV at 32 °C for 1 h, washed, and cultured at 35 °C for 4 h. Cells were fixed and immunostained with monoclonal anti-VSV-G and polyclonal anti-GM130 antibodies, followed by Alexa Fluor-conjugated anti-IgG secondary antibodies. Cells were viewed with green excitation/emission settings to detect VSV-G and red excitation/emission settings to detect GM130. Scale bar 50 μm. B, the normalized fluorescent density of VSV-G in the Golgi was measured and expressed as fold change over WT. Each bar indicates the mean ± SD of 22 to 37 cells; ∗∗∗∗p < 0.0001 versus WT and KO by one-way ANOVA, Tukey’s post-hoc test. C, primary hippocampal neurons from WT, CJD, and FFI mice were infected with VSV at 32 °C for 45 min, washed and cultured at 35 °C for 2 h. Cells were fixed and immunostained with monoclonal anti-VSV-G and polyclonal anti-giantin antibodies, followed by Alexa Fluor-conjugated anti-IgG secondary antibodies. Cells were viewed with green excitation/emission settings to detect VSV-G and red excitation/emission settings to detect giantin. Scale bar 10 μm.