Journal: Breast Cancer Research : BCR

Article Title: STAT5 is activated in macrophages by breast cancer cell-derived factors and regulates macrophage function in the tumor microenvironment

doi: 10.1186/s13058-021-01481-0

Figure Lengend Snippet: Tumor cell-derived GM-CSF activates STAT5 in macrophages. A qRT-PCR analysis for GM-CSF in TNBC (Hs578T and MDA-MB-231), ER + (MCF7) and HER2 + (BT-474) human breast cancer cells relative to expression in MCF-10A cells. B ELISA analysis for GM-CSF in CM collected from MCF-10A, MDA-MB-231, Hs578T, MCF7, and BT-474 cells. C ELISA analysis for GM-CSF in CM collected from 4T1 cells and B/B-stimulated HC11, HC11/R1, and HC11/R1-LM cells relative to EtOH controls. D Immunoblot analysis for pSTAT5, TSTAT5, and β-tubulin in BMDMs treated with No CM, tumor CM (4T1 or HC11/R1 BB), or tumor CM incubated for 1 h at 37 °C with 2.5 µg/mL neutralizing GM-CSF antibody (⍺GM-CSF Ab)

Article Snippet: STAT5 fl/fl DCM and STAT5 cKO DCM was subjected to mouse Type I Collagen ELISA (Novus Biologicals).

Techniques: Derivative Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Incubation

Journal: Breast Cancer Research : BCR

Article Title: STAT5 is activated in macrophages by breast cancer cell-derived factors and regulates macrophage function in the tumor microenvironment

doi: 10.1186/s13058-021-01481-0

Figure Lengend Snippet: STAT5 deletion in macrophages enhances tumor-promoting phenotype and impacts tumor cell migration and metastasis. A qRT-PCR analysis of genes of interest from RNA-seq associated with tumor-promoting pathways in rmGM-CSF or 4T1 CM-treated STAT5 fl/fl (blue) and STAT5 cKO (red) BMDMs. Unpaired t-test was used for statistical analysis. B Mouse Type 1 Collagen ELISA in STAT5 fl/fl unstimulated or 4T1 CM-stimulated STAT5 fl/fl and STAT5 cKO macrophage double CM (DCM). Data were analyzed using one-way ANOVA and Tukey’s multiple comparison test. C Representative immunoblot of pFAK, total FAK (FAK), and β-tubulin in 4T1 cells cultured alone or co-cultured with STAT5 fl/fl or STAT5 cKO BMDMs for 4 h. Densitometry analysis relative to loading control. D Migration analysis of 4T1 cells cultured alone or co-cultured with STAT5 fl/fl or STAT5 cKO BMDMs after 20 h. Cell counts relative to 4T1 alone in triplicate. E Kaplan–Meier curves of 4T1 cells co-injected with either STAT5 fl/fl (n = 8) or STAT5 cKO (n = 7) BMDMs in WT BALB/c mice. % Survival on Y-axes indicates proportion of mice reaching tumor size endpoint of 1cm 3 . F Quantified metastasis in H&E-stained lung sections. Lungs were sectioned at 3 different depths per mouse and analyzed for percent metastatic area per tissue section. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Scale bar: 50 μm

Article Snippet: STAT5 fl/fl DCM and STAT5 cKO DCM was subjected to mouse Type I Collagen ELISA (Novus Biologicals).

Techniques: Migration, Quantitative RT-PCR, RNA Sequencing, Enzyme-linked Immunosorbent Assay, Comparison, Western Blot, Cell Culture, Control, Injection, Staining

Journal: Inflammation Research

Article Title: Homer1 ameliorates ischemic stroke by inhibiting necroptosis-induced neuronal damage and neuroinflammation

doi: 10.1007/s00011-023-01824-x

Figure Lengend Snippet: Homer1-KD aggravated proinflammatory cytokine production. A Representative experimental diagram of chip slide carrier from ischemic penumbra cortex tissue at 8 h after pMCAO. B Heat map of protein chip results. C ELISA of TNFα in supernatants of primary neurons in different groups. D ELISA of TNFR I in supernatants of primary neurons in different groups. E ELISA of IL-1β in supernatants of primary neurons in different groups. F ELISA of Fas Ligand in supernatants of primary neurons in different groups. For in vitro cell experiments C – F , detection time is at 4 h after OGD. For C – F : ** P < 0.01, **** P < 0.0001 by one-way ANOVA analysis. Data are presented as the mean ± SD; n = 3/group. All data are representative of three independent experiments

Article Snippet: The following ELISA kits were used for detection: Mouse IL-1β ELISA Kit (ab197742; Abcam, UK), Mouse TNFα ELISA Kit (ab108910; Abcam, UK), Mouse TNFR I ELISA Kit (ab202408; Abcam, UK) , Mouse Fas Ligand ELISA Kit (ab270894; Abcam, UK).

Techniques: Enzyme-linked Immunosorbent Assay, In Vitro

Journal: Inflammation Research

Article Title: Homer1 ameliorates ischemic stroke by inhibiting necroptosis-induced neuronal damage and neuroinflammation

doi: 10.1007/s00011-023-01824-x

Figure Lengend Snippet: Homer1 protein ameliorated the pathological indexes of pMCAO. A Genotype identification of Homer1 flox/flox / Nestin-Cre +/− mice. B Representative photographs of Tunel staining of brain tissue in each group. C Quantification of result in B . D Representative photographs of p-MLKL-positive neurons of brain tissue in each group. E Quantification of result in D . F Representative experimental diagram of chip slide carrier. G IgG staining for blood–brain barrier damage. H ELISA of TNFα of ischemic penumbra cortex in different groups. I ELISA of TNFR I of ischemic penumbra cortex in different groups. J ELISA of IL-1β of ischemic penumbra cortex in different groups. K ELISA of Fas ligand of ischemic penumbra cortex in different groups. All brain tissues in the experiment were taken from ischemic penumbra cortex tissue at 8 h after pMCAO. For C , E , H – K : * P < 0.05, ** P < 0.01, and *** P < 0.001 by Student’s t test. Data are presented as the mean ± SD; n = 6/group. All data are representative of three independent experiments

Article Snippet: The following ELISA kits were used for detection: Mouse IL-1β ELISA Kit (ab197742; Abcam, UK), Mouse TNFα ELISA Kit (ab108910; Abcam, UK), Mouse TNFR I ELISA Kit (ab202408; Abcam, UK) , Mouse Fas Ligand ELISA Kit (ab270894; Abcam, UK).

Techniques: TUNEL Assay, Staining, Enzyme-linked Immunosorbent Assay