Fig. 6

Validation of PrP and XPO5 interaction. A Endogenous PrP (bait) was immunoprecipitated from a human neuroblastoma cell line (SH-SY5Y) and exportin-5 (prey) was probed by western blotting in unstressed cells (control: C), stressed cells (stress: S) and negative control (NC) IgG precipitates. B Reverse CO-IP: endogenous XPO5 (bait) was immunoprecipitated and probed with PrP (prey) by western blotting. C Abundance of XPO5 in HeLa and SH-SY5Y cells under control (cont.) and stress (str.) condition by immunoblotting. *We are not sure about the identity of these bands. D Fractionation in SH-SY5Y cells under control and stress-induced condition. W: whole cell fraction, C: cytoplasmic fraction, N: nuclear fraction. E and F Densitometry analysis of XPO5 in HeLa cells (Welch’s t-test p-value = 0.0052) and SH-SY5Y cells (Welch’s t-test p-value = 0.0004). G Densitometry analysis of all fractions in control and stress conditions. H Localization of endogenous PrP and XPO5 in stress and control (cont.) HeLa cells using confocal microscopy. Counter-staining was performed to visualize cell nuclei, scale bar = 50 μm