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: PLOS One

Article Title: MMP-2 associated imbalance of VEGF/Endostatin is linked to suppression of the PI3K/AKT/HIF-1α pathway in steroid-induced osteonecrosis of femoral head

doi: 10.1371/journal.pone.0346880

Figure Lengend Snippet: (A) Representative immunofluorescence images showing VEGF expression in the femoral head of control and SONFH groups (scale bar = 100 μm); (B) Quantification of VEGF fluorescence intensity in control and SONFH groups; (C) Representative immunofluorescence images showing Endostatin expression in the femoral head of control and SONFH groups (scale bar = 100 μm); (D) Quantification of Endostatin fluorescence intensity in these two groups; (E) Representative Western blotting images of MMP-2 expression in the femoral head of control and SONFH groups; (F) Quantification of MMP-2 expression in these two groups. (*P < 0.05, **P < 0.01).

Article Snippet: Mice in the SONFH+MMP-2 group received subcutaneous injections of recombinant MMP-2 (10 μg/kg; MedChemExpress, NJ, USA) every three days [ , ], while those in the SONFH+Endostatin group were administered daily subcutaneous injections of recombinant Endostatin (2 mg/kg; MedChemExpress, NJ, USA) [ ].

Techniques: Immunofluorescence, Expressing, Control, Fluorescence, Western Blot

Journal: PLOS One

Article Title: MMP-2 associated imbalance of VEGF/Endostatin is linked to suppression of the PI3K/AKT/HIF-1α pathway in steroid-induced osteonecrosis of femoral head

doi: 10.1371/journal.pone.0346880

Figure Lengend Snippet: (A) Representative HE staining images of the femoral head in control, SONFH, and SONFH+Endostatin groups (scale bar = 100 μm); (B) Quantitative analysis of percentage of empty osteocyte from HE staining; (C) Representative micro-CT vascular reconstructions of the femoral head in each group; (D) Calcein staining images showing mineral apposition in each group (scale bar = 25 μm); (E) Quantitative analysis of vascular volume fraction from micro-CT angiography; (F) Quantification of mineral apposition rate (MAR) in each group. (*P < 0.05, ***P < 0.001, ****P < 0.0001).

Article Snippet: Mice in the SONFH+MMP-2 group received subcutaneous injections of recombinant MMP-2 (10 μg/kg; MedChemExpress, NJ, USA) every three days [ , ], while those in the SONFH+Endostatin group were administered daily subcutaneous injections of recombinant Endostatin (2 mg/kg; MedChemExpress, NJ, USA) [ ].

Techniques: Staining, Control, Micro-CT

Journal: PLOS One

Article Title: MMP-2 associated imbalance of VEGF/Endostatin is linked to suppression of the PI3K/AKT/HIF-1α pathway in steroid-induced osteonecrosis of femoral head

doi: 10.1371/journal.pone.0346880

Figure Lengend Snippet: (A) Relative mRNA levels of Endostatin in the femoral head tissues of each group; (B) Western blotting images showing Endostatin protein expression; (C) Quantification of Endostatin protein levels; (D) Representative immunofluorescence images of Endostatin expression; (E) Quantitative analysis of Endostatin immunofluorescence intensity. (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: Mice in the SONFH+MMP-2 group received subcutaneous injections of recombinant MMP-2 (10 μg/kg; MedChemExpress, NJ, USA) every three days [ , ], while those in the SONFH+Endostatin group were administered daily subcutaneous injections of recombinant Endostatin (2 mg/kg; MedChemExpress, NJ, USA) [ ].

Techniques: Western Blot, Expressing, Immunofluorescence

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: 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: Scientific Reports

Article Title: Enhanced antiviral and antifibrotic effects of short hairpin RNAs targeting HBV and TGF-β in HBV-persistent mice

doi: 10.1038/s41598-017-04170-1

Figure Lengend Snippet: AAV-shRNA treatment decreases the expression of liver fibrosis biomarkers in HBV-persistent mice. Mice received AAV2-shRNA treatment, and serum levels of TGF-β ( a ), collagen I ( b ), and collagen III ( c ) were measured by ELISA during a 6 month period. ( d – g ) Reverse transcription quantitative PCR measurements of Tgf-β1 ( d ), Col I ( e ), Col III ( f ), and α-SMA ( g ) mRNA levels in the liver. ( h ) Western blot analysis of α-SMA in liver samples. Statistical analyses were performed using a two-way analysis of variance. P < 0.05: significant difference. Ns: the difference was not significant. Data represent the mean ± SD (n = 4).

Article Snippet: The levels of TGF-β1, collagen I, and collagen III in the liver and serum samples were determined using commercially available ELISA kits including Mouse TGF-β1 Quantikine ELISA kit, Mouse Collagen type I (Col I) ELISA kit, and Mouse Collagen type III (Col III) ELISA kit (R&D Systems, Minneapolis, MN).

Techniques: shRNA, Expressing, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot