Anti g9a 3306

Western blot analysis was carried out as previously described
[21 (link)] and using the following primary antibodies: anti-G9a (3306, Cell Signaling Technology), phospho-Akt1 (sc-33437, Santa Cruz), Akt1 (sc-8312, Santa Cruz), phospho-ERK1/2 (sc-16982, Santa Cruz), ERK1 (sc-93, Santa Cruz), E-cadherin (ab1416, Abcam), N-cadherin (610921, BD Transduction Laboratory), and α-tubulin (T5168, Sigma-Aldrich).

A Western blot analysis was carried out as previously described [31 (link)]. Briefly, HCC cells were harvested with radioimmunoprecipitation assay (RIPA) buffer (Merck Millipore, Billerica, MA, USA) containing a protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA) and phosphatase inhibitor cocktail (Merck Millipore). Protein lysates were separated via sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride membranes and immunoblotted with the following primary antibodies: anti-G9a (3306, Cell Signaling Technology, Danvers, MA, USA) and α-Tubulin (T5168, Sigma-Aldrich), as well as horseradish peroxidase-conjugated secondary antibodies. Blots were developed with enhanced chemiluminescence (ECL) Western blotting reagents (Merck Millipore).

About 200 mouse oocytes or zygotes or 30 μg testis protein per sample were mixed with SDS sample buffer and boiled for 5 min at 100 °C for SDS-PAGE. Western blotting was performed as described previously44 (link) using the antibody dilution anti-G9a (3306S, Cell Signaling Technology) at 1:1000; anti-GLP (ab41969, Abcam) at 1: 1000; anti-Actb (BS6007, Bioward) at 1: 1000; anti-Gapdh (MB001, Bioward) at 1: 1000. The membranes were subsequently incubated with HRP-conjugated secondary antibodies (1:2000; ZB2301 and ZB2305, Zhongshan Golden Bridge Biotechnology) for 1 h at 37 °C. Protein bands were detected using Thermo Supersignal West Pico chemiluminescent substrate.