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. Author manuscript; available in PMC: 2010 Jun 1.
Published in final edited form as: Biochim Biophys Acta. 2009 Mar 10;1792(6):555–563. doi: 10.1016/j.bbadis.2009.02.014

Fig. 4. More severe toxicity associated with homozygous transgenic mice (tTA +/−, pBI-cyPrP-Gal+/+).

Fig. 4

(A) Homozygous (+/+) and heterozygous (+/−) transgenic littermates at 4 weeks of age. The transgene expression was never suppressed in these mice and they were always kept in the same cage since birth. (B) Brain images of an induced homozygous transgenic mouse (Tg (+/+)) and its single transgenic littermate (control) sacrificed at 6 months of age. Bars represent 1 mm. (C) CyPrP in the forebrain of induced hetero- (+/−) or homozygous (+/+) mice were detected by immunoblot analysis with the 3F4 antibody. Equal loading was verified by re-probing the same blot with an antibody against β-actin. (D) Serial dilutions of forebrain lysates of an induced homozygous transgenic mouse and a knock-in mouse in which endogenous PrP was replaced by mouse PrP with the 3F4 epitope. The total protein concentration was indicated and PrP was detected by immunoblot analysis with the 3F4 antibody. Arrow points at cyPrP. (E) Forebrain lysates of homozygous transgenic mice were separated into supernatant (sup) and pellet (ppt) fractions by a 1-hour 100,000g centrifugation. The presence of cyPrP was determined by immunoblot analysis with the 3F4 antibody.