Figure 5. Examples of regulated expression of RIPK1 in mutant cells.
(A) Elevated expression of RIK1 in the mutant clones. RNA was isolated from mutant clones 2B2.1, 2B1.4 and 2B3, and from the parental HEK293ZeoTK cells, and analyzed by RT-PCR with RIPK1-specific primers (upper panel). The bottom panel presents results of RT-PCR on the same samples with primers for a housekeeping gene (GAPDH). (B) Dependence of RIPK1 expression in the mutant clones on the presence of the insert. Mutant clones 2B1.2 and 2B1.4 were infected with a Cre-expressing construct or the respective empty vector control. The cell lysates were compared by Western blotting with polyclonal anti-RIPK1. (C) Dependence of RIPK1 expression in the mutant clones on the function of the inserted promoter. Mutant clone 2B1.4 infected with a tTS-expressing construct was cultured with or without doxycycline. The cell lysates were analyzed by Western blotting with polyclonal anti-RIPK1 and compared to those of the parental HEK293ZeoTK cells.