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. Author manuscript; available in PMC: 2019 May 12.
Published in final edited form as: Nat Cell Biol. 2018 Nov 12;20(12):1389–1399. doi: 10.1038/s41556-018-0229-6

Figure 3. Inhibition of TBK1/IKKε sensitises cells to TNF-induced RIPK1 activity-dependent cell death downstream of LUBAC.

Figure 3

(a) MEF TNF-/- and (b) L929 cells were treated with TNF (500 ng/mL and 50 ng/mL, respectively) in the presence or absence of MRT and Nec-1s. (c) MEF TNF-/- cells of the indicated genotype were stimulated with TNF (500 ng/mL) for 6h. Loss of cell viability was determined using the Cell Titer Glo (CTG) assay. Mean +/- SEM of n=3 independent experiments. Lysates of untreated cells were analysed by Western blot. Unprocessed original scans of blots are shown in Supplementary Figure 7. (d-h) MEFs of the indicated genotype were treated with TNF (500 ng/mL) in the presence or absence of the indicated compounds. (a, b and d-h) Cell death was measured in function of time by SytoxGreen positivity. The RFU mean of 4 technical replicates of one representative experiment out of three independent experiments is represented. Representative images of indicated measurements are depicted with corresponding percentage of dead cells. Cell counting was performed manually using ImageJ. White bar in microscopy images equals 200 µm. Raw data are provided in Supplementary table 1.