Hello, everyone. I would like ask one question.

I transfected fibroblasts with EF1A-myod-puro using lipofectamine 3000. Promoter for puro is CMV. After transfection 48 hours, I did puromycin selection assay by the concentration I have chosen from killing assay using non-transfected fibroblasts. After treatment of puromycin 3 days, all my cells with myoD transfection were dead. While the cells without myoD transfection, most of them were alive even with the same treatment of puromycin (for this conditon, I also treated the cells with lipofectamine 3000, while no P3000). If the problem is myoD plasmids, I also set up another condition, with myoD plasmid transfection without puromycin treatment, all the cells were alive.

It is really strange, could someone kindly give me some suggestions?

Many thanks,

Zhou!