I am finding it difficult to make stable cells expressing GFP LC3 ( U-87 MG , IMR-32). After 24 hrs of transfection 80% of cells tend to lose expression. And within the next 24 hours, 95% cells lose expression. If I trypsinise the cells to seed in a 60 mm dish, all cells will lose expression while GFP cells are fine.
Isabelle Coupienne
You should consider transducing your cells with a retroviral vector (you can get the pBabe-puro from addgene) that's what we did for U87 (as well as for other cell lines) and as long as you keep puro in the medium, the expression is well-maintained.
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