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: BMC Cancer

Article Title: Vincristine impairs musculoskeletal development in pediatric mice

doi: 10.1186/s12885-025-15262-x

Figure Lengend Snippet: Vincristine affects trabecular, cortical bone and increases a plasma marker associated with bone resorption (CTX-1). A Quantification of trabecular bone volume fraction (Tb.BV/TV; %), B trabecular thickness (Tb.Th; μm), C trabecular number (Tb.N; μm), D trabecular spacing (Tb.Sp; μm), E connectivity density (Conn.Dn; 1/mm3), F cortical thickness (Ct.Th: mm), G cortical area (Ct.Ar; mm2), H fraction of cortical bone relative to total bone (Ct.Ar/Tt.Ar; %) in the femur of mice. I resistance to force (Polar MOI; J:mm4). J Plasma C-terminal telopeptide of type I collagen (CTX-1; pg/mL). K Representative 3-D images of trabecular bone and cortical bone. Data reported as means ± SD. Different letters denote significant differences: p <0.05. Control ( n =5); Vincristine ( n =5)

Article Snippet: The levels of circulating C-terminal telopeptide of type I collagen (CTX-1) were measured in the plasma from EDTA-treated blood collected from vehicle- and vincristine-treated mice using a specific ELISA kit (NB030329; Novus Biologicals Mouse CTX-1 ELISA Kit, Centennial, USA) according to the manufacturer’s protocol.

Techniques: Clinical Proteomics, Marker, Control

Journal: Molecular Metabolism

Article Title: Dysregulation of the Pdx1/Ovol2/Zeb2 axis in dedifferentiated β-cells triggers the induction of genes associated with epithelial–mesenchymal transition in diabetes

doi: 10.1016/j.molmet.2021.101248

Figure Lengend Snippet: Loss of β-cell identity is associated with alteration of islet microenvironment and TGFβ-dependent fibrosis . (A) Western blotting for phospho-Ser423/425 and γ-tubulin in islets from 12w Tg7 mice. (B) mRNA levels for TGFβ target genes in islets from 12w Tg7 and Wt mice obtained by qPCR (n = 6). (C) Fluorescence microscopy on pancreatic sections from Wt β−Tom and Tg7 β−Tom mice Red: tdTomato autofluorescence associated with β-cells. Scale bar: 40 μm. (D) Electron microscopy of 12w Wt and Tg7 islet preparations. Arrowhead indicates insulin granules. Arrows indicate the intercellular space between surrounding β-cells in Tg7 compared to control islets. Scale bar: 1 μm. Also shown is a zoom of the intercellular space between islet β-cells. (E) mRNA levels for ECM remodellers and collagen genes in islets from 12w Tg7 and Wt mice obtained by qPCR (n = 6). (F) Sirius red staining of pancreatic sections from 12w Tg7 mice and controls. (G) Collagen I content in islets from 12w Tg7 and Wt mice measured by ELISA. Islets from Tg7 mice were pre-treated with TGFβRI inhibitor Alk5i (1 uM) (n = 3–6). In panel G, one-way ANOVA with Sidak's multiple comparison test; otherwise, unpaired Student's t-test. Data are means ± SEM. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: The supernatant was collected, kept on ice, and processed using a Mouse Collagen Type I (COL1) ELISA kit (Abbexa, Cambridge, UK) according to the manufacturer's instructions.

Techniques: Western Blot, Fluorescence, Microscopy, Electron Microscopy, Control, Staining, Enzyme-linked Immunosorbent Assay, Comparison