Extended Data Fig. 1: DAPK3 is a positive regulator of STING signaling and some TLR pathways.
From: The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway
a, Schematic representation of the RNAi screen for IRF3 nuclear translocation. b, Immunostaining of IRF3 in HUVEC stimulated with poly (dA:dT) (1 μg/ml) (right panel) for 3 h. Scale bar, 100 μm. c, d, qRT-PCR of DAPK1/Dapk1, DAPK2/Dapk2, and DAPK3/Dapk3 in (c) human and (d) mouse cell lines. #, Not detected. e, Images of IRF3 localization in L929-mRuby-hIRF3 stimulated with poly (dA;dT) (1 μg/ml) for 3 h. Scale bar, 100 μm. f, g, (f) qRT-PCR of IFNB1 and (g) immunoblot of HUVEC transfected with indicated siRNA. h, i, (h) qRT-PCR of Ifnb1 and (i) immunoblot of BMDM transfected with indicated siRNA. (f, h) Cells were stimulated with poly (dA:dT) (0.2 μg/ml) for 4 h. j, k, (j) qRT-PCR of Il6 in L929-mRuby-hIRF3 and (k) IL6 inTHP1-Blue ISG transduced with indicated shRNA and stimulated as indicated in Fig. 1g-l. l, m, qRT-PCR of (l) Ifnb1 in L929-mRuby-hIRF3 and (m) IFNB1 inTHP1-Blue ISG transduced with indicated shRNA and transfected with poly (I:C)(LMW) and poly (I:C)(HMW)(0.1 μg/ml for L929-mRuby-hIRF3, 0.5 μg/ml for THP1-Blue ISG). n, o, qRT-PCR of (n) IFNB1 and (o) IL6 in THP1-Blue ISG transduced with indicated shRNA and stimulated with FSL-1(100 ng/ml), naked poly (I:C)(LMW)(10 μg/ml), or LPS(100 ng/ml) for 4 h. p, q, Immunoblot of (p) THP1-Blue ISG and (q) L929-mRuby-hIRF3 transduced with indicated shRNA. Data in (b-e, g, i, p, q) are representative or (f, h, j-o) the mean of three independent experiments. Values represent mean ± s.d. *P < 0.05, **P < 0.01, and ***P < 0.001. Statistical comparisons were conducted using two-tailed t-test (f, h, j-o).
