Fig. 5

Generation and analysis of transgenic offspring. a Litter size and number of transgenic positive pups in F1 generation of each litter sired by T-BST-75-GCT mice. A significant (p < 0.0001) increase in litter size of subsequent F1 generations was observed with same T-BST-75-GCT males (n = 10). b PCR screening for transgene positive pups in a litter sired by T-BST-75-GCT mice. Lanes 1, 3, and 5 shows the presence of transgene in gDNA of pups. M denotes NEB 1 kb DNA ladder; NT denotes no template control, +Ve denotes plasmid positive, 1–5 denotes lanes with gDNA of pups; Wt denotes gDNA of wild-type mice. c Slot blot for confirmation of transgene in PCR positive pups. Six out of eight (slot 1–8) showed strong positive hybridization for transgene. Wt denotes gDNA of wild-type mice, +Ve denotes plasmid positive. d Western blot of total protein isolated from liver of transgene positive pups. All the transgenic pups showed GFP expression in the liver samples as compared to wild-type. β-actin was used as internal control. e Immunostaining showing GFP expression in the liver sections of transgenic pups. i and ii show liver sections of transgenic pups. iii and iv show liver sections of wild-type mice. Scale bar: 100 μm