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. Author manuscript; available in PMC: 2013 Apr 1.
Published in final edited form as: Cancer Res. 2012 Aug 21;72(19):5060–5068. doi: 10.1158/0008-5472.CAN-12-1821

Figure 1. Men1 recombinant adenoviral vector.

Figure 1

A, Genomic organization of the Men1 recombinant vector generated by using the replication-deficient adenovirus serotype 5 (Men1.rAd5). The adenovirus has the E1 and E3 regions deleted, thereby making it replication deficient (Ad5ΔE1/ΔE3). The expression cassette consisting of 1833bp Men1 cDNA, CMV promoter and polyA+ sequence, was inserted into the E1 region of the adenoviral genome. The locations of the left and right inverted terminal repeats (LITR and RITR, respectively) together with the PacI restriction endonuclease sites are shown. A recombinant adenoviral vector expressing green fluorescent protein (GFP.rAd5) (not shown) was used as a control. B, Detection of menin and GFP expression by Western blot analysis of lysates from Men1−/− MEFs, following 48-hours of infection with either the Men1.rAd5 or GFP.rAd5 vectors or treatment with control PBS. Menin (67kDa) and GFP (27kDa) protein expression were detected only in lysates from Men1.rAd5 or GFP.rAd5-treated MEFs, respectively. Detection of α-tubulin (55kDa) expression was used as a positive control. C, Adenovirus infectivity test. Men1+/− pituitary tumour cells in culture were infected with GFP.rAd5. (i) Uninfected cells viewed by phase contrast microscopy; (ii-iv) Cells infected with 10, 102 and 103 viral particles (vp) per cultured cell, respectively, and observed by fluorescence microscopy, 24-hours after infection.