Fig 1: Exosomes derived from T cells exposed to aPD1 (Exo/aPD1) promote a phenotype of cisplatin-induced apoptosis in A549 cells via miR-4315.Cisplatin-induced cell death measure, PARP and Caspase-3 cleavages were applied to show that exosomes derived from T cells exposed to aPD1 (Exo/aPD1) promote a phenotype of cisplatin-induced apoptosis. RT-qPCR and in-cell ELIZA were applied to show that this phenomenon is associated with the miR-4315-mediated down-regulation of Bim.
Fig 2: Combined treatment with TNF-a and Ni induces apoptosis in non-small cell lung cancer cell lines. Levels of cleaved PARP and cleaved caspase-3 in the non-small cell lung cancer cell lines H292 and H1975 treated with TNF-a with or without Ni were measured by ELISA. Cleaved PARP levels were significantly increased in the (A) H292 and (B) H1975 cells treated with Ni (0.5 mM) and TNF-a (40 ng/ml). Similarly, cleaved caspase-3 levels were significantly increased in (C) H292 and (D) H1975 cells following Ni and TNF-a treatment. **P<0.01, ***P<0.001 and ****P<0.0001. Ni, nimotuzumab; TNF, tumor necrosis factor; PARP, poly ADP ribose polymerase; OD, optical density; Ctrl, control.
Fig 3: Apoptotic effect and hepatocytotoxicity of DES. (A) PC9 cells cultured with DES, Eact and Ani9 at the indicated concentrations for 24 h, with subsequent assessment of caspase-3 activity (mean ± S.D., n = 5). (B) The cells were cultured with the indicated concentrations of compounds for 24 h, and then cleaved PARP-1 activity was measured (mean ± S.D., n = 5). (C) DES concentration-dependent relative viability of HepG2 cells (red) was measured by quantifying resorufin derived from resazurin by intracellular reductases. Control experiments (Eact and Ani9) are marked for comparative analysis (gray) (mean ± S.D., n = 8). ** p < 0.01, *** p < 0.001.
Fig 4: ABT263 abrogates anti-PD1/exomiR-4315-induced resistance to chemotherapy in an in vivo model of lung cancer.A Cisplatin-induced cell death measure and PARP and Caspase-3 cleavage studies were applied to show that the phenotype of cisplatin resistance induced by exosomes derived from T cells exposed to aPD1 (Exo/aPD1) was abrogated by the use of ABT263.A Cell. B Schematic representation of our in vivo experimentations. C Graph represents the impact of treatment on tumor volume, Bim expression at mRNA (RT-qPCR experiments) and protein (ELIZA, Bim ELIZA Kit MyBioSOURCE#MBS9500064, USA) levels and on the level of serum cytochrome c (Cytochrom C ELIZA kit, Biovision#E4286-100, France). Each treatment included four mice. D Correlation between the impact of treatment on tumor volume and the level of serum cytochrome c in mice.
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