Journal: Oncotarget

Article Title: Polarization of macrophages in the tumor microenvironment is influenced by EGFR signaling within colon cancer cells

doi: 10.18632/oncotarget.12207

Figure Lengend Snippet: Cetuximab modulates macrophage polarization in an AOM/DSS mouse model A. Establishment of the AOM/DSS mouse model. AOM was injected intraperitoneally at 12.5 mg/kg body weight. After one week, mice were given drinking water containing 2.5% DSS for 5 days followed by 16 days of regular drinking water. After two cycles of DSS treatment, cetuximab (1 mg/mouse, twice a week) was injected intraperitoneally for a month, and the mice were then sacrificed. B. Representative images of colon tumors in normal (right), AOM/DSS (2AD) (middle), and cetuximab-treated AOM/DSS mice (2AD+cetu) (left). C. Tumor quantification. Cetuximab treatment (2AD + cetu) reduced tumor numbers compared to 2AD mice. D. p-EGFR (Y1068), EGFR, Arg1, and iNOS protein levels in normal mice, 2AD and 2AD+cetu mouse tumors were detected by Western blot. All experiments were repeated three times. E. Representative photomicrographs of immunostaining for p-EGFR (Y1068), PCNA, F4/80, Arg1, and iNOS. Scale bars: 25 μm. F. M1 marker (IL-12, iNOS) and M2 marker (IL-4, IL-10, Arg1) mRNA levels in normal mouse colon tissues, 2AD and 2AD+cetu mouse tumor tissues were evaluated by q-PCR. G. Percentages of CD11b + /F4/80 + and F4/80 + /CD206 + cells in normal, 2AD, and 2AD+cetu mice colon tissues were detected by flow cytometry. Colon tissues were cut into small pieces (1-2 mm) and incubated with collagenase D (1- mg/mL), dispase II (1 mg/mL), and DNase I (100 μg/mL) for 30-45 min in a shaking incubator at 37°C, and single-cell suspensions were then incubated with antibodies. Bars represent means ± SD (n = 3) for each treatment. * p

Article Snippet: Briefly, colon tissues were cut into small pieces (1-2 mm) and incubated in 10 mL PRMI 1640 medium with 10 mM HEPES and 5% fetal bovine serum containing 10 mg (1 mg/mL) collagenase D (Sigma-Aldrich), 10 mg (1 mg/mL) dispase II (Roche, Germany), and 100 μL 10 mg/ml DNase I (100 μg/mL) (Sigma-Aldrich) for 30-45 min in a shaking incubator at 37°C.

Techniques: Injection, Mouse Assay, Western Blot, Immunostaining, Marker, Polymerase Chain Reaction, Flow Cytometry, Cytometry, Incubation

Journal: The American Journal of Pathology

Article Title: Liver Repopulation and Correction of Metabolic Liver Disease by Transplanted Adult Mouse Pancreatic Cells

doi:

Figure Lengend Snippet: Enriched pancreatic ducts and pancreatic duct epithelium in culture. A: Enriched pancreatic ducts after partial digestion with collagenase D and filtration through 85-μm nylon mesh. Phase contrast, original magnification, ×100. B: Primary culture of duct cells embedded in rat-tail collagen gel with a nylon mesh. Expanding duct epithelium can be seen in the lattice. Original magnification, ×100. C: Electron microscopy of the cultured pancreatic duct cells. Their epithelial nature was apparent by the prominent microvillus processes along portions of the plasma membrane ( thick white arrows ) and formed tight junctions ( solid black arrows ) between the cohered cells. No secretary granules are seen to suggest acinar cell derivation.

Article Snippet: The harvested pancreas was immediately cut into small pieces and digested by collagenase D (Roche, Indianapolis, IN) (2.5 mg/ml, dissolved in Earle’s basic salt solution) for 25 minutes at 37°C with agitation by a magnetic stir bar.

Techniques: Filtration, Electron Microscopy, Cell Culture