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. 2006 Sep 27;26(39):9881–9891. doi: 10.1523/JNEUROSCI.2246-06.2006

Figure 4.

Figure 4.

Prmp is a highly conserved detergent-insoluble transmembrane protein. A, Amino acid alignment of rat, mouse, and human orthologs of Prmp. Prmp is a predicted single pass transmembrane protein with an N-terminal extracellular domain containing a predicted signal peptide (yellow) and a region of homology with E-cadherin (red). The predicted transmembrane domain is highlighted in blue. The C terminus contains a proline-rich domain highlighted in green. COS7 cells transfected with C-terminal Myc-tagged Prmp were fixed and permeabilized (B) or surface-stained live with anti-Myc (C). C-terminally tagged Prmp was only detected after permeabilization. DAPI staining of nuclei is blue. D, RT-PCR was performed on various adult mouse tissues. Prmp was detected in the brain, skeletal muscle, and Schwann cells. MWM, Molecular weight marker; NC, negative control; RT, no reverse transcriptase control. E, HEK293 cells were transfected with an EGFP-tagged Prmp expression vector, and 24 h after transfection, lysates were prepared with a 1% Triton X-100 extraction buffer at 4°C. Both the full-length Prmp-wt and Prmp-ct deletion mutant were found predominantly in the detergent-insoluble fraction (P). Only a small percentage of the transfected full-length Prmp-wt or Prmp-ct was recovered in the detergent-soluble fraction (S).