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: International journal of cosmetic science

Article Title: Moisturizing and antioxidant factors of skin barrier restoring cream with shea butter, silkflo and vitamin E in human keratinocyte cells.

doi: 10.1111/ics.13014

Figure Lengend Snippet: FIGURE 5 Effect of MZ on procollagen type 1 level. Values are expressed as average of triplicate experiment and are represented as mean ± SEM. * represent significant difference from H2O2 induced group (p < 0.05), # represent significant difference from untreated cells.

Article Snippet: Estimation of type I procollagen The level of type I procollagen was measured using a human type I procollagen ELISA kit (Elabscience Biotechnology, USA).

Techniques:

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