Adv cmv egfp

ADV-CMV-OLIGO2-mCherry, ADV-CMV-FOXF2- mCherry, ADV-CMV-STAT1-V5, and ADV-CMV-EGFP were produced by SignaGen Laboratories using the human cDNA sequence. Viral transduction was performed at DIV5 with a multiplicity of infection (MOI) of 20. The virus was added in fresh medium, and the medium was changed 18 hr later. Cells were harvested or fixed 96 hr after addition of virus.

For the ADV transduction experiments human iPSC- derived NSCs were plated at 700,000 cells per well of a six-well plate in 2 ml of NPM supplemented with 25 ng/ml βFGF (Peprotech, 100-18B) and 25 ng/ml activin A (Peprotech, 120–14P). Two day after plating, cells were transduced at MOI of 10 with ADV-CMV-FOXF2-mCherry (SignaGen Laboratories, SL100701), ADV-CMV-OLIG2-mCherry (SignaGen Laboratories, SL100756) and ADV-CMV-STAT1-6xHN (SignaGen Laboratories, SL110858) and MOI of 0.75 with ADV-CMV-EGFP (SignaGen Laboratories, SL100708) in 1 ml of NPM without penicillin/streptomycin. Then, the cells were placed on an orbital shaker in 37°C for 1 hr. A complete media change was performed 24 hr post-transduction. Non-transduced NSCs and NSCs transduced with ADV-EGFP were used as controls. Cells were harvested 4 days after transduction for gene expression and immunolabeling assays. ADV transduction in a human iPSC-derived NSC culture was performed twice, with three replicates each.

Adenovirus (ADV)-CMV-Nr4a1-eGFP and ADV-CMV-eGFP were produced by SignaGen Laboratories. Nr4a1-eGFP ADV was produced using the human Nr4a1 cDNA sequence. Viral transduction was performed after cells had attached for 48 h with a multiplicity of infection (MOI) of 20. The virus was added in fresh medium, and the medium was changed 18 h later. Cells were harvested or fixed 96 h following the addition of virus. ADV transduction in primary neurons was performed in four independent sets of cultures.