LTR. GFP. CMV/TRE. PGK. Blast. LTR. Tet-inducible 3T3-L1 cells. RHKD lentivirus infection at <10%. Day 1. Seeding at 1000~2000 cells per well in 96-well plate, and induction with Dox. Day 2. Day 4. Induction of differentiation. Clone screening (total ~ 450,000). Day 10-12.
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Tet-inducible 3T3-L1 cells
RHKD lentivirus infection at <10%
Seeding at 1000~2000 cells per well in 96-well plate, and induction with Dox
Induction of differentiation
Clone screening (total ~ 450,000)
Clone pickup and grow up with blasticidin, but without Dox
Re-differentiation with and without Dox