Figure 1.
Systematic Identification of Kinase Inhibitors that Modulate Naive-Primed Pluripotent Transition
(A) mESCs cultured in LIF/FBS transitioning between naive (green) and primed (red) pluripotent states.
(B) mESCs were treated with the indicated concentrations of Jak inhibitors (ruxolitinib and tofacitinib), Fgfr inhibitors (PD173074/AZD4547), or Mek1/2 inhibitors (PD0325901/PD184352). Klf4, Nanog, Dnmt3b, and Erk1/2 levels were determined by immunoblotting.
(C) 228 potent and selective kinase inhibitors were screened at 1 μM for effects on pluripotency signature. Nanog and Dnmt3b expression was determined for each inhibitor and images overlaid. Selected positive control inhibitors are highlighted.
(D) The Nanog:Dnmt3b ratio for each kinase inhibitor was determined and inhibitors ranked accordingly. Inhibitors found to alter Nanog:Dnmt3b beyond a 2-fold threshold were identified as drivers of naive or primed pluripotency. Selected positive control inhibitors are highlighted.