1. Nuclear c-Myc: A Molecular Marker for Early Stage Pemphigus Vulgaris
Lina Williamson, Thomas Hunziker, Maja M. Suter, Eliane J. Müller 
Journal of Investigative Dermatology 
Volume 127, Issue 6, Pages (June 2007)
DOI: /sj.jid
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
High c-Myc is a molecular marker for PV. Sections of paraffin-embedded biopsies of patients performed for clinical purposes were used according to the rules of the Medical Faculty. The diagnosis relied on routine clinical and histological examination in autoimmune bullous diseases (PV, PF, BP, epidermolysis bullosa acquisita, and LAD) supplemented by immunohistological (DIF) and serological (IIF) criteria. The numbers of patients analyzed are listed in Table 1. Control biopsies were obtained from two healthy donors. All biopsies were fixed with 4% formaldehyde, paraffin-embedded and serial sections of the same biopsy were used for immunofluorescence analyses using antibodies to Ki67 and c-Myc as described previously (Williamson et al., 2006). Briefly, sections were deparaffinized and antigens retrieved by microwaving three times 5minutes at 720 watt in 0.01m sodium citrate buffer pH 6.0. Antibodies used were c-Myc (1:250; Upstate Biotechnology, Lake Placid, NY; #06–340) and Ki67 (1:40; Zymed, San Francisco, CA). Nuclei were counterstained with Hoechst (H-1398, Molecular Probes, Eugene, OR). Staining was evaluated on matching biopsies and sites. One biopsy is presented with average staining (left panels) and the one with the strongest c-Myc staining (right panels). PF=pemphigus vulgaris, BP=bullous pemphigoid, TEN=toxic epidermal necrolysis, EBA=epidermolysis bullosa acquisita, AD=atopic dermatitis, SD=seborrheic dermatitis. Biopsies were processed concurrently to obtain semiquantitative results. Bars=200μm. Note that nuclear c-Myc levels were highest in PV and as reported were characteristically seen in all layers of the epidermis (Williamson et al., 2006) whereas pemphigus vulgaris antibodies were restricted to the lower part of the epidermis (inset; PG=plakoglobin; DIF=direct immunofluorescence, described in (Williamson et al., 2006); dotted line, basement membrane; discontinuous line, stratum corneum). Chosen for comparison were two different PV patients and two different sites; the PV/PF patient described in Figure 2 (peri-lesional) and a patient described previously (non-lesional; Williamson et al., 2006). Sections were processed together and experimental procedures and photographic exposures held constant to obtain semiquantitative results as described previously (Caldelari et al., 2001).
Journal of Investigative Dermatology  , DOI: ( /sj.jid )
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
C-Myc expression corresponds to disease activity in PV. In a patient with coexisting PV/PF (Muller et al., 2002), hematoxylin and eosin stained sections of formalin-fixed paraffin-embedded biopsies taken over time were evaluated by routine histology to detect blister formation (left panel). Tissue sections were examined by immunofluorescence microscopy using antibodies to Ki67 and c-Myc. Numbers indicate the increase of Ki67-positive cells in the epidermis (counted over the entire biopsy except in the lesion) as compared to normal skin (set as 1 for the control). Glucocorticosteroid (CS) treatment is indicated on the right. Biopsies were processed concurrently to obtain semiquantitative results. Bar=200μm. Note that c-Myc levels are high during active disease.
Journal of Investigative Dermatology  , DOI: ( /sj.jid )
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions