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XB-IMG-153180

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Fig. 5 – Identification of the region within Xenopus APP cDNA responsible for its aberrant promoter activity. (A) 5′-end deletion mutants of the APP-GFP construct were used in transgenesis and the generated animals were screened for fluorescence (fluo) in the brain (+: fluorescence, −: no fluorescence). The linear fragments depicted were produced via digestions with the indicated restriction enzymes. The region within APP cDNA encoding the transmembrane domain is indicated by a dashed box. pA: poly-adenylation signal. (B) Sequence analysis of the 5′-RACE PCR products to determine the start site of the aberrant transcript (indicated by an asterisk). The sequence of the nested adapter oligo used to perform the 5′-RACE PCR is underlined. (C) Western blot analysis of brain lysates of wild-type tadpoles (lane 1; background bands), tadpoles transgenic for 5′-deleted (microinjected with NaeI/PauI digested APP-GFP construct) ΔAPP-GFP (lane 2; presence of a 32-kDa product not present in lane 1) or animals expressing GFP alone (lane 3; 30-kDa GFP-product). Proteins were separated on a 12.5% SDS-PAGE gel and an anti-GFP antibody was used. Molecular weight markers (Mr) are indicated on the left.

Image published in: Collin RW and Martens GJ (2006)

Copyright © 2006. Image reproduced with permission of the Publisher, Elsevier B. V.

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