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. 2019 Mar 7;116(13):6435–6440. doi: 10.1073/pnas.1816626116

Fig. 2.

Fig. 2.

A metabolite of RIPGBM induces apoptosis in GBM CSCs by interacting with RIPK2. (A) Orbitrap MS-based metabolite identification studies in GBM-1 (GBM CSC) or primary HLF cells incubated with RIPGBM (1 μM) for 0, 12, 24, or 48 h. (B) Structure of the cyclized RIPGBM metabolite cRIPGBM generated in GBM CSCs. (C) Cell survival curves for GBM CSCs (GBM-1), human NPCs, primary human astrocyte cells, and HLFs treated with cRIPGBM for 48 h. (D) Structure of PAP reagent cRIPGBM-PAP. (E) In vitro binding of cRIPGBM-PAP to recombinant human full-length RIPK2 protein in the presence or absence of competition using underivatized cRIPGBM or RIPGBM. (F) Domain structure of RIPK2 and in vitro binding of cRIPGBM-PAP to recombinant full-length, truncated kinase domain, or truncated CARD domain human RIPK2 protein. (G) cRIPGBM-induced apoptosis in GBM-1 GBM CSCs following shRNA-mediated RIPK2 gene knockdown. Values shown are mean ± SD (*P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001).