Figure 9
From: The G127V variant of the prion protein interferes with dimer formation in vitro but not in cellulo
V127PrP forms homodimers and heterodimers with G127PrP under physiological conditions. (a) The serine variants of G127PrP and V127PrP were modified with an HA (G127PrP-HA; V127PrP-HA) or a V5 tag (G127PrP-V5; V127PrP-V5). The tags were inserted after aa 35. HeLa cells were transiently transfected with the indicated constructs. Transfected cells were lysed and HA-tagged PrP was immunoprecipitated in the absence of reducing agents with HA-agarose beads. The immunopellet was dissolved in Laemmli sample buffer containing β-mercaptoethanol and analyzed by Western blotting using an anti-V5 antibody. Western blot analysis of the input controls is shown below. Asterisk: signal corresponds to primary antibody used in the IP. (b) Quantification of the dimer formation measured densitometrically, as ratio of the dimer fraction to the HA-tagged PrP input control. WT/WT homodimers ratio was set as 1, ns not significant. Data represent mean ± SD of 3 independent experiments.
