Total protein was extracted from cells using a Protein Extraction Kit (KaiJi, Tianjin, China) according to the manufacturer's instructions. The concentration of protein was quantified via bicinchoninic acid (
BCA) protein assay (Beyotime, Shanghai, China) according to the manufacturer's instructions. Equal amounts of total protein from each sample (50 μg) were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a nitrocellulose membrane. The membranes were incubated with
rabbit anti-E-cadherin (1:1,000; Cell Signaling Technology, Boston, MA, USA),
rabbit anti-HBc (1:1,000; Dako, Glostrup, Denmark),
rabbit anti-NTCP (1:1,000; Sigma-Aldrich), rabbit anti-Na
+-K
+-ATPase (1:1,000; Abgent, San Diego, CA, USA), and
anti-actin (1:1;000; Boster, Wuhan, China) primary antibodies at 4°C overnight, and subsequently incubated with
horseradish peroxidase (HRP)-conjugated secondary antibodies (1:10,000; Boster) at 37°C for 1 h. Blots were developed using an enhanced chemiluminescence reagent (Pierce, Rockford, IL, USA) according to the manufacturer's instructions. The gray value of blots was calculated by the Image Lab.
Hu Q., Zhang F., Duan L., Wang B., Ye Y., Li P., Li D., Yang S., Zhou L, & Chen W. (2020). E-cadherin Plays a Role in Hepatitis B Virus Entry Through Affecting Glycosylated Sodium-Taurocholate Cotransporting Polypeptide Distribution. Frontiers in Cellular and Infection Microbiology, 10, 74.
