Journal: bioRxiv

Article Title: Extracellular Matrix Abnormalities Contribute to Cardiac Insulin Resistance and Associated Dysfunction in Diet-induced Obese Mice

doi: 10.1101/2023.11.14.567128

Figure Lengend Snippet: PEGPH20 treatment ameliorated high fat (HF) diet-induced myocardial fibrosis and hypertrophy. C57BL/6 mice were fed either a chow diet or a 60% HF diet for 16 weeks. After 12 weeks of HF feeding, HF-fed mice received either vehicle or PEGPH20, once every 3 days for 24 days. (A-C) Interstitial and perivascular collagens were detected by Sirius Red staining in left ventricle sections. Expression of α-SMA (smooth muscle actin) was detected by immunohistochemistry. Data were quantified by ImageJ. N=4-6. (D-G) Cardiomyocyte size was determined by Wheat Germ Agglutinin (WGA) staining. Capillary density was assessed by isolectin staining. DAPI was used to stain cell nuclei. Representative images were shown at 200x magnification (3.10 pixels/μm). Images were quantified by ImageJ. N=4-6. * p <0.05, ** p <0.01, *** p <0.005, and **** p <0.001 compared with Lean-Control; Δ p <0.05 compared with HF-Vehicle.

Article Snippet: Cardiomyocyte area and capillaries were co-stained with wheat-germ agglutinin (WGA, 2BSCIENTIFIC RL-1022-5) and isolectin B4 (Vector B-1205, 2BSCIENTIFIC B-1205-05).

Techniques: Staining, Expressing, Immunohistochemistry, Control

Journal: bioRxiv

Article Title: Extracellular Matrix Abnormalities Contribute to Cardiac Insulin Resistance and Associated Dysfunction in Diet-induced Obese Mice

doi: 10.1101/2023.11.14.567128

Figure Lengend Snippet: Pirfenidone reduced interstitial collagen deposition in the left ventricle of obese mice. C57BL/6 mice were fed with a 60% high fat (HF) diet for 16 weeks. After 12 weeks of HF feeding, mice received twice-daily treatments of vehicle or pirfenidone for 21 days. (A-C) Interstitial and perivascular collagens were detected by Sirius Red staining in left ventricle sections. Expression of α-SMA (smooth muscle actin) was detected by immunohistochemistry. Data were quantified by ImageJ. N=5-6. (D-G) Cardiomyocyte size was determined by Wheat Germ Agglutinin (WGA) staining. Capillary density was assessed by isolectin staining. DAPI was used to stain cell nuclei. N=4-5. Representative images were shown at 200x magnification (3.10 pixels/μm). Images were quantified by ImageJ. * p <0.05.

Article Snippet: Cardiomyocyte area and capillaries were co-stained with wheat-germ agglutinin (WGA, 2BSCIENTIFIC RL-1022-5) and isolectin B4 (Vector B-1205, 2BSCIENTIFIC B-1205-05).

Techniques: Staining, Expressing, Immunohistochemistry

Journal: eLife

Article Title: Kindlin-1 regulates IL-6 secretion and modulates the immune environment in breast cancer models

doi: 10.7554/elife.85739

Figure Lengend Snippet: Figure 7. Loss of tumor-derived IL-6 drives changes in Treg numbers and function but is not sufficient to reverse clearance of Kin1-NULL tumors. (A) Met-1 Kin1-NULL IL-6-CTRL or Kin1-NULL IL-6-NULL tumors were established via subcutaneous injection in FVB mice, and harvested at day 10 for immunophenotyping by flow cytometry. Gating of CD4+ T cell populations was conducted and quantified as percentage of total (alive) cells. (B) As in A with quantification of expression of markers of degranulation (CD107a) and cytotoxicity (Granzyme B) in tumor infiltrating CD8+ T cells. Example contour plots (left) and quantification of double positive cells (right). (C) Weights of tumors from mice shown in A-B. (D) Met-1 Kin1-NULL IL-6-CTRL or Kin1-NULL IL-6-NULL tumors were established via subcutaneous injection in FVB mice. Tumor size was recorded. (E) Met-1 Kin1-NULL tumors were established via subcutaneous injection in FVB mice, with 20 μg anti-IL-6 neutralising antibody administered on Day –1, 0, 4, 8, 12, and 16 post tumor cell injection. Tumor size was recorded. (F) Met-1 tumors were established in mice pre-treated with anti-CD25 to deplete Tregs. Tumor growth for individual mice (left and middle) and averages (right) are shown. Depletion demonstrated in Figure 7—figure supplement 1 (G) Tumor size at Day 17 from F. n=3–7 per group, error bars = SD. Unpaired t-test with * =< 0.05, ** =< 0.01, *** =< 0.001, **** =< 0.0001.

Article Snippet: For CD25+ cell depletion anti- mouse CD25 depleting antibody (BioXCell, via 2BScientific BP0012) and isotype control (BioXCell, via 2BScientific BP0290) were dosed at 250 μg on days −5,–4, and –3 before 1x106 cells were injected as above on day 0.

Techniques: Derivative Assay, Injection, Flow Cytometry, Expressing