Fig. 4
From: An improved Tet-on system in microRNA overexpression and CRISPR/Cas9-mediated gene editing
Development of an inducible Cas9 stable cell line. a HEK293A cells infected with lentiviruses carrying inducible FLAG-Cas9 cassette were treated with Dox for 48 h and then selected with puromycin for 5–7 d. Single cells were picked up and cultured sequentially in 96-, 24-, and 6-well plates with puromycin for several days. Positive Cas9 cell line was determined by anti-FLAG tag western blotting analysis. b The FLAG-Cas9 protein levels of the stable cell line under different Dox concentrations were assayed by western blotting. Beta-tubulin served as a loading control