Figure 2 | Cell Death & Disease

Figure 2

From: The protein level and transcription activity of activating transcription factor 1 is regulated by prolyl isomerase Pin1 in nasopharyngeal carcinoma progression

Figure 2

Pin1 interacts with ATF1 exogenously and endogenously in NPC, which is conferred by ATF1 phosphorylation sites at Thr184. (a) Mammalian two-hybrid assay showed the interaction between ATF1 and different domains of Pin1 using HEK-293 transfected with various plasmids for 24 h. (b) Coimmunoprecipitation experiments demonstrated a cellular interaction of Pin1 and ATF1 endogenously in NPC CNE2 cells. (c) Sequence logo showed the predicted phosphorylation sites at ATF1 that may affect the interaction between Pin1 and ATF1. (d) Coimmunoprecipitation experiments indicated that ATF1 Ser63Ala or Thr184Ala abolished the interaction between Pin1 and ATF1 after introducing recombinant ATF1 mutant proteins and Pin1 into CNE1 cells for 24 h. The remaining band of ATF1 was the endogenous interaction between Pin1 and ATF1 in the CNE1 cells as indicated in the first lane 'vector'. (e) Coimmunoprecipitation experiments with anti-His-tag antibody in IB to show ATF1 Ser63Ala or Thr184Ala abolished the interaction between Pin1 and ATF1 using CNE1 cells transfected with various plasmids for 24 hours. (f) Coimmunoprecipitation experiment demonstrated that phosphorylation of ATF1 at Thr184 was in the Pin1 immunoprecipitate endogenously in NPC CNE2 cells. (g) A modified ELISA assay demonstrated that the specific binding of Pin1 to phosphorylated T184 peptides in vitro. *P<0.05

Back to article page