Journal: Cancer Science

Article Title: Newcastle disease virus co‐expressing interleukin 7 and interleukin 15 modified tumor cells as a vaccine for cancer immunotherapy

doi: 10.1111/cas.13468

Figure Lengend Snippet: Construction of recombinant virus. A, Schematic representation of recombinant LX construct. The coding sequence interleukin 15 ( IL 15) and IL 7 was inserted into the sequence between P and M genes of LX genome. B, Multistep growth curve for LX and LX / IL (15+7). The virus titers were determined in triplicate by TCID 50 in DF ‐1 cells at 0, 8, 16, 24, 32 and 40 h post‐infection. The data shown are representative of the 3 experiments

Article Snippet: The expression s of IL7 and IL15 in the supernatants were measured using ELISA as described by mouse IL7 DuoSet ELISA and IL15 DuoSet ELISA (R&D Systems, Minneapolis, MN USA), respectively.

Techniques: Recombinant, Virus, Construct, Sequencing, Infection

Journal: Cancer Science

Article Title: Newcastle disease virus co‐expressing interleukin 7 and interleukin 15 modified tumor cells as a vaccine for cancer immunotherapy

doi: 10.1111/cas.13468

Figure Lengend Snippet: Expression of the interleukin 15 ( IL 15) and IL 7 in tumor cells infected with LX / IL (15+7). A, Irradiated B16 were incubated with LX / IL (15+7) or LX / RFP (100 HAU virus per 10 6 cells). After incubation for 24, 48, 72 and 96 h, the supernatants were collected and analyzed for the production of IL 15 and IL 7 (A) or IL 15‐2A‐ IL 7 (B) by ELISA

Article Snippet: The expression s of IL7 and IL15 in the supernatants were measured using ELISA as described by mouse IL7 DuoSet ELISA and IL15 DuoSet ELISA (R&D Systems, Minneapolis, MN USA), respectively.

Techniques: Expressing, Infection, Irradiation, Incubation, Virus, Enzyme-linked Immunosorbent Assay

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: BAG2 Inhibits Cervical Cancer Progression by Modulating Type I Interferon Signaling through Stabilizing STING.

doi: 10.1002/advs.202414637

Figure Lengend Snippet: Figure 6. BAG2 positively regulates STING-mediated type I interferon responses. A) RNA seq data of SiHa cells after BAG2 overexpression were analyzed by gene set enrichment analysis (GSEA), and Hallmark gene sets were used as annotated gene sets. B) Gene heat map of Hallmark Interferon 𝛼Response gene set after BAG2 overexpression in SiHa cells. C) SiHa cells were transfected with vector or GFP-BAG2 for 48 h, and then they were treated with HT- DNA for 0, 3, 6 h or cGAMP for 0, 2, 4 h. Cells were collected for detection of STING, p-TBK1, TBK1, p-IRF3, and IRF3 protein levels by Western blot. D) Vector or BAG2 plasmid was transfected in SiHa cells for 48 h, cell supernatants were collected by HT-DNA treatment for 6 hrs, and IFN-𝛽and IL7 production was measured by ELISA. E) Vector or BAG2 plasmid was transfected in U14 cells for 48 h, cell supernatants were collected by HT-DNA treatment for 6 h, and IFN-𝛽and IL7 production was measured by ELISA. F) SiHa cells and G) U14 cells were transfected with Vector or BAG2 for 48 h, then cGAMP was added for 4 h. qRT-PCR was used to measure the mRNA expression of ISG15, IFN-𝛽, MX1, and CXCL10. Data are presented as mean values ± SD, with n = 3 (B, D-G) biological independent experiments. Statistical significance was determined by two-tailed unpaired Student’s t-test (D-G).

Article Snippet: ELISA Assay: After transfecting the cells, the supernatant was collected, centrifuged to remove the precipitate, and the samples were processed according to the instructions of IL7 kit (EK207, EK107, MultiSciences Biotech) and IFN-β kit (EK2236, EK1236,MultiSciences Biotech), and then the absorbance value was detected by a microplate reader.

Techniques: RNA Sequencing, Over Expression, Transfection, Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Two Tailed Test