Immunohistochemistry was performed using EnVision+ Kits (Dako, Carpinteria, CA), according to the manufacturer’s instructions. Anti–β-catenin monoclonal antibody clone 14 (BD Transduction Laboratories, San Jose, CA) and anti-CIC rabbit polyclonal antibodies (Sigma-Aldrich Japan) were used to detect β-catenin and Capicua expression in tissues. Nuclear staining intensities to anti-CIC antibodies were evaluated in each sample and defined as CIC high (similar or higher nuclear staining intensities compared with those of adjacent nontumor hepatocytes) or low, respectively. Immunofluorescence analysis was performed using anti-CIC rabbit polyclonal antibodies (Sigma-Aldrich Japan) and Alexa 488 fluorescein isothiocyanate–conjugated anti-rabbit IgG as primary and secondary antibodies, respectively. All images were obtained using a Biorevo BZ-9000 Fluorescence Microscope system (Keyence, Osaka, Japan).
Hashiba T., Yamashita T., Okada H., Nio K., Hayashi T., Asahina Y., Hayashi T., Terashima T., Iida N., Takatori H., Shimakami T., Kawaguchi K., Arai K., Sakai Y., Yamashita T., Mizukoshi E., Takamura H., Ohta T., Honda M, & Kaneko S. (2020). Inactivation of Transcriptional Repressor Capicua Confers Sorafenib Resistance in Human Hepatocellular Carcinoma. Cellular and Molecular Gastroenterology and Hepatology, 10(2), 269-285.
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