Journal: The FASEB Journal

Article Title: Obesity reduces mammary epithelial cell TGFβ1 activity through macrophage‐mediated extracellular matrix remodeling

doi: 10.1096/fj.202000228rr

Figure Lengend Snippet: FIGURE 4 Obesity increases macrophages expressing decorin around mammary ducts and within breast lobules. A, Fold change of decorin expression relative to cyclophilin or HPGRT of ASCs, mammary epithelial cells (MEC) and macrophages isolated from mammary glands of CD or HFD-fed FVB/N mice (n = 6 mice/group). B, Number of F4/80+ macrophages surrounding mouse mammary ducts divided by the ductal area (n = 6 FVB/N mice/group). C, Number of CD68+ macrophages within the lobular and ductal area in human breast tissue (n = 6 breast samples/ group). Data analyzed with Student's t test, mean ± SEM. Magnification bar = 50 µm

Article Snippet: Tissue sections were stained for immunohistochemistry and immunofluorescence using SMAD4 (1:100, PA534806, Thermo Fisher Scientific), phosphorylated SMAD2 (pSMAD2, 1:250, 44-244G, Thermo Fisher Scientific), decorin (1:25, sc-22613, Santa Cruz Biotechnologies, Dallas, TX, USA), F4/80 (1:250, BM8, 12302, BioLegend, San Diego, CA, USA), CD68 (1:200, 50-112-2648, Thermo Fisher Scientific), alpha-smooth muscle actin (SMA, 1:5000, 5528, MilliporeSigma), and TGFβ1 (1:250, LS-B14345, LSBio, Seattle, WA, USA) as described.8 Sections were imaged using Nikon Eclipse 80 t microscope with NIS Elements BR 3.2 software.

Techniques: Expressing, Isolation

Journal: The FASEB Journal

Article Title: Obesity reduces mammary epithelial cell TGFβ1 activity through macrophage‐mediated extracellular matrix remodeling

doi: 10.1096/fj.202000228rr

Figure Lengend Snippet: FIGURE 5 Macrophage depletion in obese mice decreases decorin expression in ECM and increases downstream targets of the TGFβ1 pathway within the epithelium. A, Schematic of macrophage depletion experiment in HFD-fed mice. B, Weight gain of female FVB/N mice fed HFD starting at 3 weeks of age. Mice received treatment with either IgG or anti-F4/80 antibodies (n = 4 mice/group, repeated measures ANOVA with Sidak's multiple comparison test). C, Number of F4/80+ macrophages surrounding mammary ducts divided by the ductal area (n = 4 mice/ group; Student's t test). D, Percent area decorin in ECM surrounding mammary ducts (n = 4 mice/group; Student's t test). E, Percent pSMAD2+ cells in mammary ducts (n = 4 mice/group; Student's t test). F, Percent SMAD4+ cells in mammary ducts (n = 4 mice/group; Student's t test). Bars represent mean ± SEM. Magnification bar = 50 µm

Article Snippet: Tissue sections were stained for immunohistochemistry and immunofluorescence using SMAD4 (1:100, PA534806, Thermo Fisher Scientific), phosphorylated SMAD2 (pSMAD2, 1:250, 44-244G, Thermo Fisher Scientific), decorin (1:25, sc-22613, Santa Cruz Biotechnologies, Dallas, TX, USA), F4/80 (1:250, BM8, 12302, BioLegend, San Diego, CA, USA), CD68 (1:200, 50-112-2648, Thermo Fisher Scientific), alpha-smooth muscle actin (SMA, 1:5000, 5528, MilliporeSigma), and TGFβ1 (1:250, LS-B14345, LSBio, Seattle, WA, USA) as described.8 Sections were imaged using Nikon Eclipse 80 t microscope with NIS Elements BR 3.2 software.

Techniques: Expressing, Comparison