Figure 4.
In vitro expression of insulin and C-peptide or proinsulin by adenovirus transduced mouse Sertoli cells. (A) mSC cultured overnight as a monolayer on six-well plates with DMEM plus 10% FBS were either nontransduced (MOI of 0; lane 2) or transduced with AdRSVGFP (lane 3), AdCMVhInsWT (lane 4), or AdCMVhInsM (lane 5) adenoviral vectors at a MOI of 100. Cells were collected after 2 days and RT-PCR was performed for insulin or β-actin. Lane 1 is the 1kb Plus DNA Ladder (Invitrogen). (B) mSC cultured overnight as a monolayer on chamber slides were transduced with AdCMVhInsM (a and d), AdCMVhInsWT (b and e), or AdRSVGFP (c and f) adenoviral vectors at a MOI of 100. Slides were collected after 2 days, fixed with 1% paraformaldehyde and immunostained for insulin (a–c) or C-peptide (d–f). All sections were counterstained with hematoxylin.