1. Paraneoplastic Pemphigus Sera React Strongly with Multiple Epitopes on the Various Regions of Envoplakin and Periplakin, Except for the C-Terminal Homologous Domain of Periplakin 
Yoshiko Nagata, Tadashi Karashima, Fiona M. Watt, Wolfgang Salmhofer, Tamotsu Kanzaki, Takashi Hashimoto 
Journal of Investigative Dermatology 
Volume 116, Issue 4, Pages (April 2001)
DOI: /j x
Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Schematic diagram for the structures of human envoplakin and periplakin and the positions of the 11 truncated recombinant GST-fusion proteins, including seven domain-specific proteins and four proteins of the junctional region. The left and right figures summarize the structures and the recombinant proteins of envoplakin and periplakin, respectively. In each figure, the upper panel depicts an entire structure of each protein. The middle panel indicates the positions and the residue numbers of all the recombinant proteins used in this study. In the lower panel, the positions of all the cDNA constructs used as templates for PCR are shown.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
By immunoblotting of normal human epidermal extracts, all the PNP sera reacted strongly with envoplakin and periplakin, whereas a few non-PNP sera reacted weakly with proteins showing the same migration. By immunoblotting of epidermal extracts, six representative PNP sera reacted with the doublet of the 210 kDa envoplakin and the 190 kDa periplakin (lanes 1–6), whereas either of these protein bands was also recognized by some particular sera from cases of BP (lanes 7–9), PV (lanes 10–11), and PF (lane 12), in addition to their specific antigens. The positions of the 250 kDa desmoplakin I (DPKI), the 210 kDa envoplakin (ENV), and the 190 kDa periplakin (PPK) are shown on the left. The positions of the 230 kDa BP230, the 180 kDa BP180, the 160 kDa Dsg1, and 130 kDa Dsg3 are shown on the right.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Multiple recombinant proteins of envoplakin were strongly recognized by the majority of the PNP sera. Panels 1–4 show the results of all the 26 PNP sera (“PNP”) and normal sera (“Cont”) on immunoblotting of ENV-N, ENV-M, ENV-C, and ENV-H, respectively. An arrowhead on the left of each panel indicates the position of intact recombinant protein.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Multiple recombinant proteins of periplakin were strongly recognized by the majority of the PNP sera, whereas the C-terminal homologous domain of periplakin reacted with only three PNP sera. Panels 1–3 show the results of all the 26 PNP sera (“PNP”) and normal sera (“Cont”) on immunoblotting of PPK-N, PPK-M, and PPK-H, respectively. An arrowhead on the left of each panel indicates the position of intact recombinant protein.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
A few PNP sera reacted with the small recombinant proteins of junctional regions between each domain of envoplakin and periplakin. Panels 1–4 show the results of immunoblotting of the recombinant proteins of junctional regions, ENV-NM, ENV-MC, PPK-NM, and PPK-MC, respectively. In each panel, lanes 1–6 show the results for PNP sera, lane 7 for anti-GST polyclonal antibody, and lanes 8–10 show the results for normal control sera. In all the panels, the lanes with the same number show the reactivity of the same serum.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

7. Figure 6
A few non-PNP sera reacted weakly with some recombinant fusion proteins, particularly with the C-terminal homologous domain of periplakin. Panels 1–7 show the results of representative sera from BP (lanes 1 and 2), PV (lanes 3 and 4), and PF (lanes 5 and 6), as well as normal sera (lanes 7 and 8), a control PNP serum (lane 9), and anti-GST polyclonal antibody (lane 10) on immunoblotting of ENV-N, ENV-M, ENV-C, ENV-H, PPK-N, PPK-M, and PPK-H, respectively. In all the panels, the lanes with the same number showed the reactivity of the same serum.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

8. Figure 7
Combination method of immunoprecipitation and immunoblotting confirmed that some non-PNP sera react with envoplakin and periplakin. The diluted epidermal extracts were first immunoprecipitated with various sera, and immunoprecipitated materials were fractionated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and blotted. The anti-envoplakin antibody (CR5) detected envoplakin in the blots for various sera, which showed the protein comigrating with envoplakin by immunoblotting (lanes 1–4). Anti-periplakin antibody (CR3) detected periplakin in the blots for various sera, which showed the protein comigrating with periplakin by immunoblotting (lanes 5–10).
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions