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Fig. 4 | Journal of Animal Science and Biotechnology

Fig. 4

From: An improved Tet-on system in microRNA overexpression and CRISPR/Cas9-mediated gene editing

Fig. 4

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

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