Fig. 3: C-terminally truncated ZBP1 induces cell death in vitro. | Cell Death & Differentiation

Fig. 3: C-terminally truncated ZBP1 induces cell death in vitro.

From: ZBP1 causes inflammation by inducing RIPK3-mediated necroptosis and RIPK1 kinase activity-independent apoptosis

Fig. 3

A Scheme depicting structure of FL-ZBP1 and ZBP1ca. B Cell death measured by Draq7 uptake in iMEFs stimulated with combinations of 1 µg doxycycline (dox), emricasan (emri), Nec1s and RIPK3 inhibitor GSK’827. Cell death was normalized to 2 h of Draq5 treatment for each genotype. Representative of at least 3 different experiments. Mean ± SEM are shown. C Immunoblot analysis of protein extracts from iMEFs transduced with doxycycline (Dox)-inducible FL-ZBP1- or ZBP1ca-expressing vectors or stimulated with IFNα. Representative of 3 different experiments. D Anti-Flag immunoprecipitates in iMEFs expressing dox-induced Flag-tagged FL-ZBP1 or Flag-tagged ZBP1ca stimulated with dox. Representative of 5 different experiments. E Immunoblot analysis of protein extracts from iMEFs transduced with Dox-inducible ZBP1ca-, FL-ZBP1 or ZBP1ca mZα-expressing vectors after treatment with increasing Dox concentrations. Representative of 3 different experiments. F Representative images of iMEFs expressing the indicated ZBP1 constructs after overnight doxycycline (+) treatment immunostained for ZBP1. G Cell death measured by Draq7 uptake in iMEFs stimulated with 8 µg dox. Representative of 3 different experiments. Mean ± SEM are shown. H Relative mRNA expression of RNA from iMEFs expressing FL-ZBP1 or ZBP1ca after indicated times of dox treatment. Graph shows mean ± SEM of three different experiments, the same LPS + TNF treated control RNA was used for all experiments.

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