Journal: Cells

Article Title: Fibronectin Regulation of Integrin B1 and SLUG in Circulating Tumor Cells

doi: 10.3390/cells8060618

Figure Lengend Snippet: Immunomodulatory mechanisms of CTCs derived from a syngeneic mouse model of HCC. ( A–D ) Major histocompatibility complex class I (MHCI) expression was assessed using flow cytometry. CTCs have decreased MHCI cell surface protein expression in comparison to primary tumor-derived cell lines. Both isotype and MHCI antibodies were conjugated to fluorescein isothiocyanate (FITC). Isotype measuring background is shown in red. MHCI signal is shown in green; N = 5. ( E ) Analysis of 111 different cytokines secreted into cell media reveals consistent and significantly decreased secretion of endostatin, CXCL5, and proliferin in CTCs in comparison to primary tumor-derived cells; N = 2. ( F – H ) Endostatin, CXCL5, and proliferin signals from cytokine array were quantified. Data are presented as mean ± standard error of the mean (SEM).

Article Snippet: Either FITC anti-mouse MHCI antibody (BD Biosciences, NJ, USA, cat#: 553565) or its corresponding FITC Mouse Isotype control (BD Biosciences, NJ, USA, cat#: 553456) was added to cells and allowed to incubate for 1 h. The samples were fixed with 2% paraformaldehyde for 30 min at 4 °C, washed, and re-suspended in 1X PBS.

Techniques: Derivative Assay, Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: Age-Related Impairment of Innate and Adaptive Immune Responses Following Intranasal Herpes Simplex Infection

doi: 10.1101/2022.01.05.475170

Figure Lengend Snippet: 6-week, 36-week and 68-week-old mice were infected intranasally with 1 × 10 6 pfu of HSV-1 strain. Mice were euthanized 2 days post infection and a single cell suspension from lungs were obtained after collagenase treatment. The lung cells were stained for epithelial cell markers, TLR-3 and activation markers and then analyzed by FACS. ( A ) Timeline of infection and immunological analyses. ( B ) Gating strategy used to characterize lung derived cells. Lymphocytes were identified by a forward scatter (FSC) and side scatter (SSC) gate. Singlets were selected by plotting forward scatter area (FSC-A) vs. forward scatter height (FSC-H). CD45 negative cells were then gated. The epithelial cell population in CD45-gated cells was defined by the EpCAM + cells. ( C ) Representative histograms and average mean fluorescence intensity (MFI) of TLR-3 expression on the surface of lung epithelial cells ( D ) ICAM expression on epithelial cells ( E ) MHCI expression on lung epithelial cells of 6-week, 36-week and 68-week-old mice at 2 days post-infection with HSV-1 (solid black) or mock-infection with DMEM (solid white). Data is representative of two separate experiments.

Article Snippet: The following antibodies were used: anti-mouse CD8 PerCP (BD Biosciences, San Jose, CA), anti-mouse CD11b FITC (BD Biosciences), anti-mouse CD103 APC (BD Biosciences) anti-mouse CD11c (BD Biosciences) anti-mouse CD45 APC-cy7 (BioLegend, San Diego, CA), anti-mouse TLR3 (BD Biosciences), anti-mouse MHCI (BD Biosciences), MHCII (eBioscience), CD4 Percp, CD107 a FITC, CD107 b FITC (BD Biosciences) and anti-mouse IFN-γ PE-cy7 (clone XMG1.2, BioLegend).

Techniques: Infection, Staining, Activation Assay, Derivative Assay, Fluorescence, Expressing