Journal: Biology of Sex Differences

Article Title: Sex differences in disease severity and immune responses in murine and human inflammatory arthritis

doi: 10.1186/s13293-026-00840-w

Figure Lengend Snippet: Male CIA mice exhibit higher disease severity compared to females. Male and female CIA and saline control mice were monitored for disease severity and assigned clinical scores from day 21 after the first CII challenge until the end of experiment (day 29). (A) Line graphs representing the mean clinical scores of mice starting from day 1 to day 29. On day 29 after the first CII challenge, mice were euthanized by cardiac puncture under anesthesia for blood and serum collection and storage at −80 °C. Serum concentrations of (B) anti-mouse collagen type II antibodies (left panel) and anti-bovine collagen type II antibodies (right panel) were analyzed by ELISA. N = 5 mice per group. One of two independent experiments. Simple linear regression analysis was performed to determine the statistical difference between the lines. One-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison post hoc test was used to determine the statistical significance between the groups. (*/ # p ≤ 0.05, ** p ≤ 0.005 and ***/ ### p ≤ 0.0005). In Fig. 1A, # significance of comparisons between CIA and saline control mice; *significance of comparisons between sexes of CIA mice

Article Snippet: Serum levels of mouse anti-collagen antibodies (autoantibodies) and bovine anti-collagen antibodies (antibodies to the immunizing antigen) were determined by ELISA using a Mouse Anti-mouse Type II Collagen IgG Antibody Assay Kit and Mouse Anti-Bovine Type II Collagen IgG Antibody Assay Kit, respectively, according to the manufacture’s protocol (Chondrex Inc. WA, USA).

Techniques: Saline, Control, Enzyme-linked Immunosorbent Assay, Comparison

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: The FASEB Journal

Article Title: Amelogenin Null Mice Develop Osteoarthritis, While Its Application Mitigates Disease Phenotypes in a Rat Model

doi: 10.1096/fj.202500827R

Figure Lengend Snippet: Immunohistochemical analyses of the knee joints from 12‐month‐old amelogenin‐null and wild‐type mice. Representative images of immunohistochemistry for type II collagen (A, C) and matrix metalloproteinase‐13 (MMP‐13) (B, D) in wild‐type (WT) (A, B) and amelogenin‐null (C, D) mice. Amelogenin‐null mice exhibited reduced type II collagen levels and increased MMP‐13 expression compared to wild‐type mice. These changes were accompanied by cartilage destruction and the presence of hypertrophic chondrocytes (black arrowheads). Enlarged pictures were taken from the regions marked with black frames. Brown staining indicates positive expression of type II collagen or MMP‐13. Scale bars: 250 μm for overall images; 62.5 μm for enlarged images.

Article Snippet: Indirect immunohistochemistry was performed using primary antibodies directed against type II collagen (AM26374PU‐N, ORIGENE) or matrix metalloproteinase 13 (MMP‐13; TA321485, ORIGENE).

Techniques: Immunohistochemical staining, Immunohistochemistry, Expressing, Staining

Journal: Oncology letters

Article Title: SMOC1 silencing suppresses the angiotensin II-induced myocardial fibrosis of mouse myocardial fibroblasts via affecting the BMP2/Smad pathway.

doi: 10.3892/ol.2018.8989

Figure Lengend Snippet: Figure 4. SMOC1 silencing mitigates the oxidative stress in MFBs induced by Ang II. ELISA was performed to evaluate the (A) MDA content, (B) LDH content and (C) SOD activity in MFBs, MFBs transfected with an empty vector, and Ang II‑induced MFBs transfected with an empty vector or si‑SMOC1. *P<0.05 and **P<0.01 vs. NC; ^P<0.05 and ^^P<0.01 vs. Ang II+NC. SMOC1, SPARC‑related modular calcium binding 1; MFBs, myocardial fibroblasts; Ang II, angiotensin II; MDA, malondialdehyde; LDH, lactate dehydrogenase; SOD, superoxide dismutase; si, small interfering RNA; NC, negative control.

Article Snippet: The expression of TGF‐β1, COL-I and COL-III were determined by ELISA kits: Mouse TGF‐β1 ELISA Kit (E‐EL‐M0051c; Elabscience, Wuhan, Hubei, China), COL1 ELISA kit (E‐EL‐M0325c; Elabscience) and COL‐III ELISA kit (E‐EL‐M0316; Elabscience).

Techniques: Enzyme-linked Immunosorbent Assay, Activity Assay, Transfection, Plasmid Preparation, Binding Assay, Small Interfering RNA, Negative Control

Journal: Oncology letters

Article Title: SMOC1 silencing suppresses the angiotensin II-induced myocardial fibrosis of mouse myocardial fibroblasts via affecting the BMP2/Smad pathway.

doi: 10.3892/ol.2018.8989

Figure Lengend Snippet: Figure 5. SMOC1 silencing downregulates the expression levels of fibrosis‑associated proteins. (A) ELISA was performed to measure the TGF‑β1, COL‑I and COL‑III expression in MFBs, MFBs transfected with an empty vector, and Ang II‑induced MFBs transfected with an empty vector or si‑SMOC1. (B) Reverse transcription‑quantitative polymerase chain reaction and (C) western blot analysis were performed on the expression levels of FN, TGF‑β1, COL‑I and COL‑III in MFBs, MFBs transfected with an empty vector, and Ang II‑induced MFBs transfected with an empty vector or si‑SMOC1. *P<0.05, **P<0.01, and ***P<0.001 vs. NC; ^P<0.05 and ^^P<0.01 vs. Ang II+NC. SMOC1, SPARC‑related modular calcium binding 1; FN, fibronectin; TGF‑β1, transforming growth factor β1; COL, collagen; Ang II, angiotensin II; MFBs, myocardial fibroblasts; si, small interfering RNA; NC, negative control.

Article Snippet: The expression of TGF‐β1, COL-I and COL-III were determined by ELISA kits: Mouse TGF‐β1 ELISA Kit (E‐EL‐M0051c; Elabscience, Wuhan, Hubei, China), COL1 ELISA kit (E‐EL‐M0325c; Elabscience) and COL‐III ELISA kit (E‐EL‐M0316; Elabscience).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation, Polymerase Chain Reaction, Western Blot, Binding Assay, Small Interfering RNA, Negative Control

Journal: PLoS ONE

Article Title: Mesenchymal Stromal Cells Improve Salivary Function and Reduce Lymphocytic Infiltrates in Mice with Sjögren's-Like Disease

doi: 10.1371/journal.pone.0038615

Figure Lengend Snippet: Photomicrographs of CD45 − /TER119 − cells that undergone chondrocytic, osteogenic and adipocytic differentiation when cultured in differentiation media. Lower right photomicrograph represents CD45 − /TER119 − cells cultured in non-differentiating media (negative control). Collagen II (red) and DAPI (blue nuclei) are used to identify chondrocytes. Von kossa (black) and oil red stainings were used to identify osteoblasts and adipocytes, respectively. Graphs represent the number, size, and frequency of colony-forming unit fibroblasts (CFU-F) of CD45 − /TER119 − cells versus whole bone marrow cells. All three graphs show that CD45 − /TER119 − cells had higher CFU-F than whole bone marrow cells (* P <0.05).

Article Snippet: Immunofluorescence staining to collagen II was used to identify chondrocytes (Collagen II antibody, # AF3615; Northern Lights 557-conjugated anti-sheep IgG, # NL010, R&D Systems, MN).

Techniques: Cell Culture, Negative Control