Journal: Oncotarget

Article Title: ST2/IL-33 signaling promotes malignant development of experimental squamous cell carcinoma by decreasing NK cells cytotoxicity and modulating the intratumoral cell infiltrate

doi: 10.18632/oncotarget.25768

Figure Lengend Snippet: The frequencies of macrophages (F4/80 + ), B (CD19 + ) and T cells (CD4 + and CD8 + ), and dendritic cells (CD11c + ) were determined using immunostaining and FACS analysis in tumor lesions that were harvested from mice on day 126. The bars show (A) the absolute number of leukocytes and the number of (B) CD4 + T lymphocytes, (C) CD8 + T lymphocytes, (D) B lymphocytes, (E) dendritic cells, (F) and macrophages present in the tumor lesions. (G) Frequencies of CD11b + (myeloid cells), CD119 + (M1) and CD124 + (M2) cells in the tumor microenvironment. The data are representative of three experiments (n = 5 per group). * P < 0.05.

Article Snippet: For immunostaining, FcγRs were blocked in mAb 2.4G2, and the cells were then stained for surface markers using PerCP-, PE- and FITC-conjugated antibodies against CD45 (30-F11), CD19 (1D3), CD3 (145-2C11), CD4 (RM4-5), CD8 (53-6.7), CD11b (M1/70), CD27 (LG3A10), DC (33D1), CD124 (mIL4R-M1), NKp46 (29A1.4), KLRG1 (2F1) (BD Biosciences), F4/80 (BM8) (eBioscience), and CD119 (REA189) (Miltenyi Biotec).

Techniques: Immunostaining

Journal: Cell reports

Article Title: Opposing tumor-cell-intrinsic and -extrinsic roles of the IRF1 transcription factor in antitumor immunity

doi: 10.1016/j.celrep.2024.114289

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: inVivo MAb anti-mouse IFNγR (CD119), clone GR-20 , BioXcell , Cat# BE0029; RRID:AB 1107576.

Techniques: Immunofluorescence, Staining, Control, Recombinant, Protease Inhibitor, Sample Prep, Purification, DNA Purification, Reverse Transcription, Proliferation Assay, BIA-KA, Transfection, Derivative Assay, Cloning, Sequencing, Plasmid Preparation, Software

Journal: PLoS ONE

Article Title: A role for the immune system-released activating agent (ISRAA) in the ontogenetic development of brain astrocytes

doi: 10.1371/journal.pone.0248455

Figure Lengend Snippet: ISRAA acted on embryonic brain astrocytes early at E7 and late at E21. It stimulated them to produce IFN-γ. E7 astrocytes did not express IFN-γR; therefore, they could not respond to the anti-proliferative effects of IFN-γ and continued to proliferate. However, E21 cells did express IFN-γR, and, accordingly, they ceased proliferation and began to differentiate.

Article Snippet: Cells were then blocked by 2% normal sheep serum diluted in PBS for 30min, followed by the addition of the rabbit anti-mouse IFN-γ receptor (IFN-γR) antibody (MyBioSource, Inc., San Diego, CA, USA) and the rabbit anti-ISRAA polyclonal antibody [ ], diluted at 1/1,000, for 24h at 4°C.

Techniques:

Journal: PLoS ONE

Article Title: A role for the immune system-released activating agent (ISRAA) in the ontogenetic development of brain astrocytes

doi: 10.1371/journal.pone.0248455

Figure Lengend Snippet: Cell Released Capturing ELISA (CRC-ELISA) was used assess spontaneous and ISRAA-induced IFN-γ production by embryonic astrocytes. E7 and E21 astrocytes spontaneously produced this cytokine. Stimulation with recombinant ISRAA significantly increased IFN-γ production by E7 and E21 (*** p <0.05). However. the induction of IFN-γ was significantly higher in E21 than in E7 astrocyte cultures (*** p <0.05). Experiments were conducted twice with alike findings. Bars represent the means ± SDs of 12 cultures.

Article Snippet: Cells were then blocked by 2% normal sheep serum diluted in PBS for 30min, followed by the addition of the rabbit anti-mouse IFN-γ receptor (IFN-γR) antibody (MyBioSource, Inc., San Diego, CA, USA) and the rabbit anti-ISRAA polyclonal antibody [ ], diluted at 1/1,000, for 24h at 4°C.

Techniques: Enzyme-linked Immunosorbent Assay, Produced, Recombinant

Journal: PLoS ONE

Article Title: A role for the immune system-released activating agent (ISRAA) in the ontogenetic development of brain astrocytes

doi: 10.1371/journal.pone.0248455

Figure Lengend Snippet: IFN-γR was detected by immunohistochemistry. Counter staining was conducted with haematoxylin. As depicted in the micrograph, E7 astrocytes did not express receptors for IFN-γ (Fig 4A), while E21 astrocytes showed strong expression of IFN-γR (Fig 4B). Pictures were acquired with light microscopy (x630). The micrograph is a representation of several fields scanned in each chamber. All fields showed expression of IFN-γR on E21, but not E7.

Article Snippet: Cells were then blocked by 2% normal sheep serum diluted in PBS for 30min, followed by the addition of the rabbit anti-mouse IFN-γ receptor (IFN-γR) antibody (MyBioSource, Inc., San Diego, CA, USA) and the rabbit anti-ISRAA polyclonal antibody [ ], diluted at 1/1,000, for 24h at 4°C.

Techniques: Immunohistochemistry, Staining, Expressing, Light Microscopy

Journal: PLoS ONE

Article Title: A role for the immune system-released activating agent (ISRAA) in the ontogenetic development of brain astrocytes

doi: 10.1371/journal.pone.0248455

Figure Lengend Snippet: The rationale behind the IFN-γ blocking experiment was to find if the ISRAA-induced proliferation suppression of E21 was achieved by IFN-γ produced in response to ISRAA or not. Incubation with anti-IFN-γ antibody in cultures of E21 astrocytes 15 min prior to ISRAA stimulation significantly inverted the inhibitory effects of ISRAA on proliferation (***p<0.05), but had no effect on E7 (bars with horizontal lines). The IFN-γ blocking effect was obvious on E21 as they express the IFN-γR, but not E7. Experiments were conducted twice with alike results. Bars represent the means ± SDs of 12 cultures.

Article Snippet: Cells were then blocked by 2% normal sheep serum diluted in PBS for 30min, followed by the addition of the rabbit anti-mouse IFN-γ receptor (IFN-γR) antibody (MyBioSource, Inc., San Diego, CA, USA) and the rabbit anti-ISRAA polyclonal antibody [ ], diluted at 1/1,000, for 24h at 4°C.

Techniques: Blocking Assay, Produced, Incubation

Journal: PLoS ONE

Article Title: A role for the immune system-released activating agent (ISRAA) in the ontogenetic development of brain astrocytes

doi: 10.1371/journal.pone.0248455

Figure Lengend Snippet: To demonstrate the IFN-γ signalling pathway activation on embryonic brain astrocytes after ISRAA stimulation, tyrosine kinase activity and nuclear localisation of the transcription factor STAT1 in E21 astrocytes were examined using Western blot analyses and immunohistochemistry respectively. A. Phosphorylation of tyrosine-specific protein. Cell lysates were prepared 5min, 15min and 60min after ISRAA stimulation. Note the slight amounts of tyrosine phosphorylated proteins in the non-stimulated control lysates (Ctrl) on lane 1. ISRAA stimulation resulted in a strong and fast rise in tyrosine phosphorylation, which reached its highest point by 15min and declined to the control level at 60min (ISRAA; lane 2, 3, 4). An irrelevant antibody (Ctrl Ab; lane 5, 6, 7) was used to control the specificity of the phosphorylated protein. The band representing activated TPKs (lane 3, 15 min on ISRAA) was not detected. Tyrphostin A47 (a tyrosine kinase specific inhibitor) blocked the ISRAA induced protein kinase as demonstrated by the inhibition of the at the various detection times (A47; lane 8, 9, 10). B. Nuclear localisation of the STAT1. Nuclear localisation of the STAT1 protein was detected in E21 astrocytes 5min, 15min and 60min after ISRAA stimulation. The shown pictures were obtained by a digital camera using a fluorescence microscope at x630 magnification after 15min incubation. B.1 represents non-stimulated cells (note the generalised cytoplasmic antigen distribution before ISRAA treatment; red stained cytoplasm and black non-stained nuclei). B.2 depicts the nuclear translocation of STAT1 15 min after ISRAA stimulation (heavily stained nuclei in red). B.3 demonstrates the blocking effects of tyrphostin A47 on the ISRAA promoted nuclear translocation of STAT1.

Article Snippet: Cells were then blocked by 2% normal sheep serum diluted in PBS for 30min, followed by the addition of the rabbit anti-mouse IFN-γ receptor (IFN-γR) antibody (MyBioSource, Inc., San Diego, CA, USA) and the rabbit anti-ISRAA polyclonal antibody [ ], diluted at 1/1,000, for 24h at 4°C.

Techniques: Activation Assay, Activity Assay, Western Blot, Immunohistochemistry, Phospho-proteomics, Control, Inhibition, Fluorescence, Microscopy, Incubation, Staining, Translocation Assay, Blocking Assay