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. 2021 Mar 1;296:100490. doi: 10.1016/j.jbc.2021.100490

Figure 1.

Figure 1

Mutant PrPs localize abnormally in HeLa cells. HeLa cells were transfected with plasmids encoding WT, PG14, CJD, or FFI PrP-EGFP fusion protein. After 48 h cells were fixed, permeabilized, stained with anti-PDI (A) or anti-GM130 (B) polyclonal antibody followed by Alexa Fluor 647 (red)-conjugated anti-rabbit IgG secondary antibody, and reacted with Hoechst 33258 (blue) to stain the nuclei. Scale bar 10 μm. C, the number of cells showing PrP-EGFP on the plasma membrane (PM) with no or little colocalization with PDI or GM130/giantin, or intense colocalization with PDI (ER) or GM130/giantin (Golgi) was counted and expressed as a percentage of the total. Each bar indicates the mean ± SD of three independent experiments evaluating a total of 210 WT, 198 PG14, 191 CJD, and 204 FFI PrP-EGFP transfected cells. ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 versus WT by two-way ANOVA, Tukey’s post-hoc test.