Journal: Science signaling

Article Title: The injury response to DNA damage in live tumor cells promotes antitumor immunity.

doi: 10.1126/scisignal.abc4764

Figure Lengend Snippet: Fig. 8. Live injured chemotherapy-treated tumor cells require DNA-damage signaling for the DC-mediated T cell IFN- response. (A) B16-Ova cells were treated with doxorubicin at 0.5, 1, 2.5, or 5 M for 24 hours, washed, and incubated with primary bone marrow–derived dendritic cells (BMDCs) for another 24 hours. After this, OT-1 CD8+ T cells were added and evaluated for intracellular IFN- 15 hours later. Quantification of IFN-+CD8+ T cells from three independent experiments is shown. Error bars represent SEM. ****P < 0.0001 by ANOVA, followed by Dunnett’s multiple comparisons test. (B) B16-Ova cells were treated with doxorubicin, etoposide, or mitoxan- trone at the doses indicated for 24 hours and the live (attached) fractions were analyzed by fluorescence microscopy after staining with DAPI (blue channel) and anti-H2AX (red channel). Representative images are shown. Scale bar, 20 m. (C) Quantification of H2AX foci intensity from the experiment in (B) is shown. n ≥ 200 cells per condition. Width in violin plot indicates frequency for each observed value from maximum to minimum, with dotted line indicating median. ****P < 0.0001 by ANOVA followed by Dunnett’s multiple comparisons test compared to the DMSO control. (D) B16-Ova cells were treated with doxorubicin, etoposide, or mitoxantrone at the doses indicated for 24 hours, followed by separation of the live and dead fractions and analysis of respective lysates by Western blotting. Blots are representative of two independent experiments. (E) Quantification of IFN-+CD8+ T cells (from three independent experiments) induced by BMDCs after incubation with B16-Ova cells that were cotreated with 50 M etoposide and 10 M ATM kinase inhibitor KU-55933, ATR kinase inhibitor AZD6738, or DNA-PK inhibitor NU7441. First lane (−) defined as in Fig. 1C. Error bars represent SEM. **P < 0.01 and ****P < 0.0001 by ANOVA, followed by Dunnett’s multiple comparisons test versus the “Etop 50 M” group.

Article Snippet: KU-55933, AZD6738, and NU7441 were from SelleckChem.

Techniques: Incubation, Derivative Assay, Fluorescence, Microscopy, Staining, Control, Western Blot

Journal: Viruses

Article Title: Proteomic Analysis of Vero Cells Infected with Pseudorabies Virus

doi: 10.3390/v14040755

Figure Lengend Snippet: Primers used in this study.

Article Snippet: The primary antibodies used in this study were specific for XRCC5 (16389-1-AP, Proteintech, Rosemont, IL, USA), XRCC6 (10723-1-AP, Proteintech, Rosemont, IL, USA), β-actin (66009-1-Ig, Proteintech, Rosemont, IL, USA), VP5 (prepared in our lab), and gB (prepared in our lab).

Techniques: Sequencing

Journal: Viruses

Article Title: Proteomic Analysis of Vero Cells Infected with Pseudorabies Virus

doi: 10.3390/v14040755

Figure Lengend Snippet: Validation of proteomics data by western blot and RT-qPCR. ( A ) Vero cells infected with PRV for 24 h were collected and western blot was performed to detect the expression of XRCC5 and XRCC6 with corresponding antibodies. ( B ) The western blot and proteomics ratio of XRCC5 and XRCC6. ( C ) Relative XRCC6 transcription in Vero cells. ( D ) Relative XRCC5 transcription in Vero cells. ( E ) The expression of XRCC5 and XRCC6 in PK-15 infected with PRV. ( F ) The expression of XRCC5 and XRCC6 in CRL-2843 infected with PRV. ** indicates significance at a 99% confidence interval ( p < 0.01).

Article Snippet: The primary antibodies used in this study were specific for XRCC5 (16389-1-AP, Proteintech, Rosemont, IL, USA), XRCC6 (10723-1-AP, Proteintech, Rosemont, IL, USA), β-actin (66009-1-Ig, Proteintech, Rosemont, IL, USA), VP5 (prepared in our lab), and gB (prepared in our lab).

Techniques: Biomarker Discovery, Western Blot, Quantitative RT-PCR, Infection, Expressing