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. Author manuscript; available in PMC: 2020 Apr 25.
Published in final edited form as: Cell Rep. 2020 Jan 21;30(3):699–713.e4. doi: 10.1016/j.celrep.2019.12.073

Figure 1. RIPK Promotes Cytokine mRNA and Protein Production in LPS-Activated MOLF Macrophages.

Figure 1.

(A and B) TNF-α and CXCL-1 mRNA (A) and protein (B) levels in unstimulated, LPS-, LZ-, LZN-, Z-, and N-stimulated B6, MOLF, RIP1 Ki, RIP3 Ki, and Mlkl−/− BMDMs.

(C) Cell death as measured by propidium iodide incorporation over 5 h in B6, MOLF, RIP1 Ki, and Mlkl−/− BMDMs.

(D) RIP1, total, and phospho-MLKL levels in B6 and MOLF BMDMs stimulated as indicated with LZ ± 5 IU IFNβ priming overnight.

(E) IFNb, ISG15, and IRF7 mRNA levels as a readout for constitutive IFN signaling in B6 and MOLF, BMDMs, I.P. macrophages, and blood.

(F and G) Total STAT1 (F) and ISG15 and IRF7 mRNA (G) levels as a readout of reconstitution of constitutive IFN signaling in B6 and MOLF BMDMs stimulated as indicated with LZ ± 5 IU IFNβ priming overnight.

In all panels, BMDMs were stimulated with LPS, LPS/zVAD (LZ), LPS/zVAD/Nec-1 s (LZNs), zVAD (Z), or Nec-1 s (Ns) as indicated. ELISA and qPCR data are shown as ±SD from three independent experiments compared using two-way ANOVA: n.s. (p > 0.05), **p < 0.01, ***p < 0.001, and ****p < 0.0001. Kinetic cell death and western blots are representative of three or more independent experiments. Kinetic data are presented as the mean ± SD of triplicate wells.

See also Figure S1.