Co-immunoprecipitation assay and western blotting (WB) were performed as previously described45 (link). Lysates were prepared from HuH-7 cells transfected with plasmids of interest 48 hr after transfection. The antibodies used in co-immunoprecipitation were anti-Cx43 antibody (Sigma), anti-T7-tag antibody (Novagen), anti-HA-tag antibody, anti-DYKDDDDK (FLAG)-tag antibody (Wako Pure Chemical), and normal mouse and normal rabbit IgG (Santa Cruz Biotechnology). The antibodies used in WB were anti-Cx43 antibody (Sigma), anti-Hsc70 antibody (StressMarq), anti-T7-tag antibody (Novagen), anti-HA-tag antibody, anti-DYKDDDDK (FLAG)-tag antibody, anti-cMyc-tag antibody (Wako Pure Chemical), and anti-GAPDH antibody (Santa Cruz Biotechnology). The signals were measured by Image Quant LAS500 (GE Healthcare UK Ltd., Buckinghamshire, England).
Hino H., Dai P., Yoshida T., Hatakeyama T., Harada Y., Otsuji E., Okuda T, & Takamatsu T. (2015). Interaction of Cx43 with Hsc70 regulates G1/S transition through CDK inhibitor p27. Scientific Reports, 5, 15365.
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