Anti spag9

IHC was carried out as previously described [55 (link), 74 (link)]. The antibodies used for IHC are as follows: anti-KSHV LANA (Advanced Biotechnologies Inc., Columbia, MD, USA), anti-SPAG9 (Abcam, Cambridge, MA, USA), anti-rabbit immunoglobulin G (IgG) (Beyotime Institute of Biotechnology, Nantong, Jiangsu, China), and anti-smooth muscle actin (SMA) rabbit polyantibody (Abbiotec, San Diego, CA, USA). Secondary antibodies used in this study were horseradish peroxidase (HRP)-labeled goat anti-rat or anti-rabbit from Beyotime Institute of Biotechnology (Nantong, Jiangsu, China). DAB (3,3'-diaminobenzidine) Peroxidase (HRP) Substrate Kit (VECTOR LABORATORIES, INC., Burlingame, USA) was used to visualize staining.

HCC samples or HCC cells were lysed with RIPA peptide lysis buffer (Beyotime Biotechnology, Jiangsu, China) containing 1 % protease inhibitors (Pierce) and Western blotting analysis were performed according to standard procedures. Primary antibodies were used as follows: anti-SPAG9 (1:1000, Abcam), anti-JNK, anti-p-JNK, anti-c-Jun, and anti-MMP9 (1:2000, Abcam) and anti-GAPDH (1:3000, Huabio). Protein bands were developed using the Enhanced Chemiluminescence (ECL) system and were visualized and quantified by using the ChemiScope Western Blot Imaging System (Clinx Science Instruments Co., Ltd).