CCl4 Treatment in mice

Liver Systems Medicine : striving to develop non-invasive methods for diagnosing and treating NAFLD by combining mathematical modeling and biological research. LiSyM, is a multidisciplinary research network, in which molecular and cell biologists, clinical researchers, pharmacologists and experts in mathematical modeling examine the liver in its entirety. LiSyM research focuses on the metabolic liver disease non-alcoholic fatty liver disease (NAFLD), which includes non-alcoholic steatohepatitis ...

In one in five people with NAFLD, the functioning liver cells, the hepatocytes, are replaced by connective tissue. Eventually this fibrosis becomes irreversible. In this state the liver is like a ‘scar that never heals’. Through it, the liver loses many of its vital functions. LiSyM-Pillar II wants to know more about which factors promote fibrosis and the conditions under which fibrosis becomes irreversible How can fibrosis be diagnosed as early as possible? Researchers in the pillar are also ...

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Supplmentary Materials, Methods, Figures and Tables

List of used antibodies

Publications in which periportal fibrosis has been reported after repeated doses of CCl4.

Materials and Methods

CYP2E1 activity in HepG2-E47 was measured. TAM inhibits (not significantly) the CYP2E1 activity compared to alcohol and DMSO-treated cells. C34 cell line was used as a negative control. Bars represent the means±SD of 3 biological replica. Student’s t test (paired t test) was used to check the statistical differences upon TAM incubation (JPG 1530 KB)

Protein level of CYP2E1 was analyzed by co-immunofluorescence staining immediately after attachment (3h) and 24 hours later. b and c) Protein and mRNA levels of CYP2E1 were analyzed upon TAM pretreatment. d and e) Protein and mRNA levels of catalase were assessed upon TAM pretreatment. Bars represent the means±SD of 3 mice (JPG 3000 KB)

Formalin-fixed livers were processed and stained with HE, CYP2E1, GS, F4/80 and CD45 to visualize and quantify hepatocellular necrosis, metabolizing enzymes, resident macrophages and recruited immune cells, respectively. Slides were scanned and the signal was randomly quantified in 15 images per liver. Yellow dashed lines indicate the necrotic areas. Yellow arrows refer to the positive signals. Scale bars are 200µm for HE and CYP2E1 and 100µm for GS, F4/80 and CD45 (JPG 6105 KB)

RNA microarray made with primary hepatocytes from female mice with hepatocyte-specific, embryonic knockout of Smoothened

This SOP - Standard Operating Procedure for Sample Collection and Preservation of Blood and Endometrial Tissue were developed at NIH University of California San Francisco Human Endometrial Tissue and DNA Bank by Elizabeth Sheldonand, Ramsey A McIntire