Journal: BMC Genomics

Article Title: Comparison of the molecular and cellular phenotypes of common mouse syngeneic models with human tumors

doi: 10.1186/s12864-019-6344-3

Figure Lengend Snippet: Immune subsets in syngeneic models. a In silico immune cell deconvolution of syngeneic tumor samples. Syngeneic models exhibited various immune cell type infiltrations with major NK cell infiltration predicted in CT26 models. b Comparison of estimated total T-cell fraction of leukocyte in selected mouse syngeneic models and their corresponding human tumors. Human data were downloaded from Gentles et al. . Total T-cell fraction plotted here is the sum of all predicted T-cell subsets including CD4+, CD8+, Treg, and gamma-delta T-cells. c CD3 staining for T-cells, CD11b staining for myeloid cells, and F4/80 staining for macrophage

Article Snippet: Sections were incubated with primary antibodies, CD11b (AbCam, EPR1334, Citrate, 0.088 μg/ml), F4/80 (Spring Bioscience, M4152, Citrate 2.5 μg/ml), RA3-6B2, Borg, 2.5 μg/ml), CD3 (Spring Bioscience, SP162, Borg, 1.33 μg/ml), vimentin (Cell Marque, SP20, Citrate, 3 μg/ml), E-cadherin (AbCam, EP700Y, Citrate, 0.18 μg/ml), for 1 h, washed in TBS and incubated with SignalStain Boost IHC Detection Reagent (Cell Signaling Technologies, Beverly, MA, USA) for 30 min.

Techniques: In Silico, Comparison, Staining

Journal: Biomedicines

Article Title: Tumors Established in a Defective Immune Environment Reprogram the Oncogenic Signaling Pathways to Escalate Tumor Antigenicity

doi: 10.3390/biomedicines12040846

Figure Lengend Snippet: Tumor regressors are edited when transplanted into BALB/c mice with the increased recruitment of tumor-infiltrating lymphocytes. ( a ) Representative immunofluorescence staining and ( b ) statistical analysis show CT26/BALB/c and CT26/SCID tumors infiltrated by CD3+ T cells on day 14; ( c ) Representative immunofluorescence staining and ( d ) statistical analysis show CT26/BALB/c and CT26/SCID tumors infiltrated by CD3+ T cells on day 21. Data are representative of 3–4 independent experiments. Bar graphs reflect mean ± SD. Statistical analyses were performed with the unpaired Student’s t test. ****, p < 0.0001; HPF, high-power field.

Article Snippet: Slides were incubated with primary antibodies: CD3 (Abcam, Cambridge, UK, clone SP162, Catalog No. ab135372, dilution factor: 1:200), CD11c (Abcam, clone EP1347Y, Catalog No. ab52632, dilution factor: 1:200), and CD103 (Abcam, clone EPR22590-27, Catalog No. ab224202, dilution factor: 1:500).

Techniques: Immunofluorescence, Staining

Journal: Nature Communications

Article Title: Post-resolution macrophages shape long-term tissue immunity and integrity in a mouse model of pneumococcal pneumonia

doi: 10.1038/s41467-024-48138-y

Figure Lengend Snippet: WT C57BL6/J mice were administered intranasal S. pneumoniae with animals ( A ) experiencing a transient weight loss. Lungs were digested and samples analysed by polychromatic flow cytometry for markers of acute inflammation including ( B ) total leucocytes, ( C ) neutrophils, ( D ) pro-inflammatory cytokines and ( E ) bacterial clearance (Colony Forming Units/mL of bronchoalveolar lavage fluid). Flow cytometry was also used to profile ( F ) total lymphoid and ( G ) myeloid cells throughout inflammation, resolution and weeks following resolution. The temporal profile of ( H ) neutrophils (GR1 + ) as well as ( I ) macrophages (F4/80 + ) and CD3 T cells (CD3 + ) were confirmed at tissue level, panels H-I. A Alveoli, B bronchiole, BV blood vessel. Sections are representative of n = 3 independent experiments. Data were analysed by one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons test. A p value of <0.05 was taken as the threshold of significance with graphical representation as; p < 0.05 = *, p < 0.01 = ** and p < 0.001 = *** and presented as mean ± SEM. ( n = 5–8 mice/group).

Article Snippet: For day 14 samples, sections were incubated with primary conjugated antibodies, Alexa Fluor® 488 anti-mouse F4/80 [BM8, Biolegend], Alexa Flour® 467 anti-mouse COX-2 [EPR3777, Abcam-Conjugated using Thermo Fisher labelling kit A20186], CF®555 anti-mouse CD3 [SP162, Abcam-Conjugated using Biotium Mix-n-Stain™ CF® Dye Antibody Labelling Kits] and primary unconjugated antibodies anti-mouse EP4/PTGER4 (4A2A12, Proteintech) anti-human mPGES-1 (Polyclonal, Cayman Chemical 160140) and anti-mouse Siglec F/CD170 (S17007L, Biolgend) overnight at 4 °C.

Techniques: Flow Cytometry