Journal: Proceedings of the National Academy of Sciences of the United States of America
Article Title: A cell type-selective apoptosis-inducing small molecule for the treatment of brain cancer
Figure Lengend Snippet: 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
Article Snippet: Cell pellets and growth media of GBM CSC and control cell types were extracted following drug treatment and subjected to quantitative high-resolution Orbitrap LC-MS analysis (Thermo Fisher Scientific).
Techniques: Mass Spectrometry, Incubation, Generated, In Vitro, Binding Assay, Recombinant, shRNA