Live imaging of prions reveals nascent PrPScin cell-surface, raft-associated amyloid strings and webs

ScGT1 cells growth-arrested by a 6d MMC/BrdU treatment were processed using the FA-denaturing protocol and co-stained with mAb SAF32 and with mAb 8H4 followed by subtype-specific secondary antibodies: cy2 anti-IgG2b for SAF32 (white) and RRX anti-IgG1 for 8H4. Results for SAF32 are shown in this stack. Cells were photographed using an Axiovert 200M microscope using a 100x NA1.3 Plan Neofluar oil objective and a Zeiss FS38HE filter cube. Thirty five images at 300nm z increments were acquired using the MetaMorph 6.2 software. The z-stack was processed by blind 3D deconvolution using the Huygens software (Scientific Volume Imaging). Twenty sequential planes from the 3D deconvolved stack are included in the data presented here. Webs of branching PrP strings cover large areas of the cell periphery; elsewhere, the paper shows that strings are found on the cell surface. Plane #19 of 20 is shown in Fig. 1D,b in the paper. Please notice that a fragment of this is also included in the paper in Fig. 2C,a. The 8H4 co-staining data are included in another z-stack under "Figure 2, C,b".