PMC:4385186 / 20955-22098 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/4385186","sourcedb":"PMC","sourceid":"4385186","source_url":"https://www.ncbi.nlm.nih.gov/pmc/4385186","text":"Mouse Xenograft Assay and Immunohistochemistry\nTo determine the effects of ZNF750 on tumorigenesis in vivo, we used a mouse xenograft assay with 12- to 14-week-old BALB/c nude female mice. We injected 2 × 106 KYSE150 cells stably depleted of ZNF750, scrambled control vector, wild-type ZNF750, or p.Ser70∗ ZNF750 into nude mice (n = 6 mice/group). The growth rates of xenograft tumors were measured for 4 weeks, after which mice were sacrificed. After 28 days, tumors were removed, snap frozen in liquid nitrogen, and stored at –80°C. Tumor size was measured with calipers. Additionally, formalin-fixed paraffin-embedded xenograft tumors were immunohistochemically stained with a monoclonal mouse Ki-67 antibody (Zhongshan). In brief, sections were incubated with the Ki67 antibody working dilution for 14 hr at 4°C and then detected with PV8000 (Zhongshan) and the DAB detection kit (Maixin), producing a dark-brown precipitate. Slides were counterstained with hematoxylin. All images were captured at 20×. The nuclear amount of Ki-67 was analyzed with Aperio Nuclear v.9 software. Statistical analyses were performed with GraphPad Prism 5.0.","divisions":[{"label":"title","span":{"begin":0,"end":46}}],"tracks":[]}