Journal: Journal of Extracellular Vesicles

Article Title: An extracellular vesicular mutant KRAS‐associated protein complex promotes lung inflammation and tumor growth

doi: 10.1002/jev2.12307

Figure Lengend Snippet: EV‐KRAS complex facilitates inflammation through the IL‐17A/FGF21 axis. (a) HE staining of lung tissue from mice (blocked with IL‐17R/Fc or anti‐FGF21 antibody) intratracheally administered extracellular vesicle (EV) KRAS complex (liposome‐EV‐KRAS). (b) ELISA for TNFα, IL‐17A and IL‐6 from lung tissue from mice (blocked with IL‐17R/Fc or anti‐FGF21 antibody) intratracheal administered liposome‐EV‐KRAS. (c) Adenosine triphosphate (ATP) cell proliferation assay showing proliferation of Jurkat cells in response to lung tumour lysate with and without IL‐17A depletion using IL‐17A immunoprecipitation. (d) Luciferase assay for FGF21 promotor assessment, TC1 cells treated with respective lung tumour lysate with and without IL‐17A depletion using IL‐17A immunoprecipitation. (e) Western blot for PI3K, SIRT1, NFκB‐ TC1 cells treated with respective lung tumour lysate with and without FGF21 depletion using FGF21 immunoprecipitation. (f) qRT‐PCR for genes involved in EMT pathway‐ TC1 cells treated with respective lung tumour lysate with and without FGF21 depletion using FGF21 immunoprecipitation. For in vivo treatment mice were administered with 50 μl of 10 × 1010 liposome particles/ml and for in vitro treatment 2 × 108 liposome particle/ml for respective group were used. Data are mean ± SEM from three replicates using one‐way ANOVA, ** p < 0.01.

Article Snippet: In vivo cytokine neutralization experiments involved administering mAbs against FGF21 (Abclonal, Woburn, MA) and recombinant Mouse IL‐17RA/IL‐17R Fc Chimera Protein (R&D System) intraperitoneally.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Proliferation Assay, Immunoprecipitation, Luciferase, Western Blot, Quantitative RT-PCR, In Vivo, In Vitro