Fig. 3: PK6 and PK68 block cellular activation of RIPK1, RIPK3, and MLKL upon necroptotic stimuli. | Cell Death & Disease

Fig. 3: PK6 and PK68 block cellular activation of RIPK1, RIPK3, and MLKL upon necroptotic stimuli.

From: Discovery of potent necroptosis inhibitors targeting RIPK1 kinase activity for the treatment of inflammatory disorder and cancer metastasis

Fig. 3

a, b Bone marrow-derived macrophages from C57BL/6 mice or rat were pretreated with DMSO, PK6(10 μM), or PK68 (100 nM) for 1 h and subsequently treated with indicated stimuli for 24–30 h. Cell viability was determined by measuring ATP levels. L, LPS (20 ng/ml); P, poly(I:C) (50 μg/ml); S (100 nM); Z (10 μM). Data are represented as the mean ± standard deviation of triplicates. c HT-29 cells were treated with the indicated compound for 1 h prior to the treatment of T (40 ng/ml), S (100 nM), and Z (20 μM) for additional 8 h. Cell lysates were harvested and subjected to western blot analysis for phosphorylation of RIPK1, RIPK3, and MLKL. d The effects of PK6 and PK68 on the formation of RIPK3 puncta. HT-29 cells stably expressing Flag-tagged RIPK3 were pretreated with indicated compounds for 1 h prior to the treatment of T (100 ng/ml), S (100 nM), and Z (20 μM) for additional 12 h. The distribution of RIPK3 was detected by immunofluorescence. *P < 0.05. **P < 0.01. ***P < 0.001

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