<p>(A) Characterization of glycomutants (PrP<sup>C</sup>G1, PrP<sup>C</sup>G2, and PrP<sup>C</sup>G3) and PrP<sup>C</sup>WT for the study by Western blot analysis, using an antibody directed against the 3F4 epitope. Clones with similar amounts of overexpressed 3F4 tagged PrP<sup>C</sup> as assessed by densitometric analysis of Western blots were used for these analyses (see graph). Relative expression of various PrP<sup>C</sup> forms is shown in percentages of PrP<sup>C</sup>WT that was set to 100%. (B) Assessment of plasma membrane (non-permeabilized) and intracellular (permeabilized) localization of PrP<sup>C</sup> glycomutants by confocal microscopy shows presence of PrP<sup>C</sup> at the plasma membrane and intracellularly (scale bar is 10 µm). (C) Assessment of DRMs localization of PrP<sup>C</sup> glycomutants by Triton X-100 extraction at 4°C and sucrose density gradient centrifugation showing correct localization of PrP<sup>C</sup> glycomutants with flotillin-positive DRM containing fractions.</p
