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. Author manuscript; available in PMC: 2008 Sep 22.
Published in final edited form as: J Invest Dermatol. 2007 Jul 26;127(12):2807–2817. doi: 10.1038/sj.jid.5700970

Figure 1.

Figure 1

The hBPAG2 cDNA transgene construct expressed protein of appropriate immunoreactivity and molecular weight in an immortalized GABEB keratinocyte line. a: An immortalized GABEB keratinocyte line transfected with the transgene construct showed bright, diffuse cytoplasmic staining for hBPAG2 in IF microscopy studies. b: In contrast, GABEB keratinocytes transfected with the same construct lacking hBPAG2 cDNA (i.e., cells transfected with a “mock” construct and subjected to selection in the same manner) showed no staining for hBPAG2. Nonimmune rabbit sera showed no reactivity to these cells (data not shown). c: Immunoblot studies of extracts from transgene- and mock-transfected GABEB keratinocytes identified hBPAG2 as a 180 kD polypeptide in the former (lane 4) but not the latter (lane 2). Immortalized GABEB keratinocytes transfected with the same hBPAG2 cDNA in an alternate vector (i.e., pcDNA3) served as an additional positive control (lane 3) as did extracts of normal HKs (lane 1). Nonimmune rabbit sera showed no reactivity to these extracts (data not shown). Ticks in the left margin correspond to 250 and 116 kD molecular weight markers; the arrow points to a 180 kD band corresponding to hBPAG2. d: Tg mice express hBPAG2 in epidermal BM. Cryosections of skin from man, a Tg mouse, and a Wt mouse (top, middle, and bottom panels, respectively) were studied by IF microscopy using rabbit antiserum developed against the carboxy terminus of hBPAG2 (Masunaga et al., 1997) (left panels) or normal rabbit serum (control, right panels). hBPAG2 was specifically expressed in the epidermal BMs of human and Tg skin (arrows, top and middle left panels); this antiserum showed no specific reactivity to the skin of a Wt mouse (bottom left panel). There was no cytosolic, suprabasal, or intradermal expression of hBPAG2 in any skin samples. Control IF microscopy studies using normal rabbit serum were negative (right panels).