Extended Data Fig. 4: The PIKfyve inhibitor YM201636 blocks STING-mediated TBK1 activation.
From: PtdIns(3,5)P2 is an endogenous ligand of STING in innate immune signalling
(a-b) Prolonged YM201636 treatment blocks HT-DNA- or cGAMP-stimulated phosphorylation of TBK1 and STING in BJ cells. BJ cells pretreated with DMSO or 5 μM of YM201636 for 18 h were transfected with HT-DNA (2 μg/ml) for 3 h (a) or delivered with 100 nM cGAMP for 1-3 h (b) and whole cell lysates were analyzed by immunoblotting. (c-d) Prolonged YM201636 treatment blocks cGAMP-stimulated TBK1 signaling in L929 (c) and MEF (d) cells. Cells pretreated with DMSO or 5 μM of YM201636 for 18 h were delivered with 100 nM cGAMP for 1 hour and whole cell lysates were analyzed by immunoblotting. (e) Prolonged YM201636 treatment does not affect SeV-stimulated TBK1 activation in BJ cells. BJ cells pretreated with DMSO or 5 μM of YM201636 for 18 h were infected with SeV for 6 h and whole cell lysates were analyzed by immunoblotting.
