Nourseothricin N-Acetyl Transferase: A Positive Selection Marker for Mammalian Cells
Figure 2
Expression of NAT allows generation of stable cell lines expressing heterologous proteins using NTC.
(A) H-PNHB cells were infected with retroviruses encoding NAT and p100HA, and 12 hours later treated with NTC for selecting stable clones. Images of the infected cells, cells after 48 hours of selection and the generated stable cell line is shown. The black arrowheads indicate dying/dead cells. (B) Expression of HA-p36 in H-NH cells was induced for 24 hours with indicated amounts of Dox in the presence and absence of NTC. (C) Expression of HA-p100 in selected H-PNHB cells was induced with Doxycycline (Dox) for 24. Whole cell lystes were prepared after 24 hours and expression of HA-p100 was detected using an anti-HA antibody. Bands corresponding to HA-p36 and HA-p100 are indicated.