For the coimmunoprecipitation experiments, HuH - 7 cells transfected with pcDNA - SRα / FLAG - Sp 1 were harvested and cell lysates were prepared using
immunoprecipitation lysis buffer [20 mmol / L Tris - Cl (pH 7.5), 150 mmol / L NaCl, 10 % glycerol, and 1 % Triton X-100].
After extensive washing with
immunoprecipitation lysis buffer, the immunoprecipitated proteins were analyzed by immunoblotting using specific antibodies against DDX3 and FLAG epitope (ab1257, Abcam, Cambridge, United Kingdom).