Verification of Cell Line Deletions

The HeLa cervical carcinoma cell line (JCRB9004) was obtained from the JCRB Cell Bank and used as a control. Huh7.5 cells (Blight et al., 2002 (link)) were kindly provided from Dr. Charles M. Rice. Huh7.5.1 cells (Zhong et al., 2005 (link)) were kindly provided from Dr. Francis V. Chisari. Genomic DNA was prepared from the cell lines using the Blood Genomic DNA Extraction Mini Kit (Favorgen, Ping-Tung, Taiwan, ROC). Deletions in the genomes were verified by PCR experiments using a previously reported procedure (Sakuma et al., 2018 (link)). Briefly, PrimeSTAR GXL DNA Polymerase (Takara Bio, Otsu, Japan) was used for amplification. The reaction mixture, containing 60 ng genome DNA, was denatured at 98°C for 1 min and then subjected to 40 cycles, consisting of 98°C for 10 s, 61°C for 15 s, and 68°C, for 1 min. The amplified products were electrophoresed on an agarose gel and visualized with the Gel Doc EZ imager (Bio-Rad, Hercules, CA, United States). Then, 1 kb Plus DNA Ladder was used as a molecular marker (Thermo Fisher Scientific, Waltham, MA, United States).