Anti mkk3

For IHC, HCC tissue microarrays analysis (TMA) slides were purchased from Alenabio (Xi'an, China). The primary antibodies used in IHC were anti-p-p38 (Cell signaling technology (CST), Boston, MA, USA, #4511), anti-p38 (Abcam, Hongkong, China, #ab31828), anti-p-MKK3/6 (CST, #9236), anti-MKK3 (Epitomics, Burlingame, CA, USA, #1728), anti-MKK6 (Epitomics, #1640), anti-YAP (CST, #12395), anti-c-Jun (CST, #2315), anti-c-Myc (Epitomics, #1472), anti-ki67 (Abcam, #ab15580), anti-AFP (Abcam, #ab46799) and anti-SIRT1 (for mSIRT1, CST, #2028; for hSIRT1, Abcam, #ab32441). For IF, the primary antibodies used were anti-p-p38 (for mp-p38, CST, #4511; for h-p-p38, Abcam, #ab45381), anti-p38 (Abcam, #ab31828), anti-p-MKK3/6 (CST, #9236) and anti-FLAG-tag (CST, #8146). For WB, the primary antibodies used were anti-GAPDH (CST, #5174), anti-p-p38 (CST, #4511), anti-p38 (Abcam, #ab31828), anti-SIRT1 (for mSIRT1, CST, #2028; for hSIRT1, Abcam, #ab32441), anti-p-MKK3/6 (CST, #9236), anti-MKK3 (Epitomics, #1728), anti-MKK6 (Epitomics, #1640), anti-FLAG-tag (CST, #8146), anti-AFP (Abcam, #ab46799) and anti-YAP (CST, #12395). All IHC, IF and WB experiments were performed conventionally and the protocols are available elsewhere.

Co-IP was performed as described previously [53 (link)]. The reagents used were: protein A/G-Sepharose (Novex, Oslo, Norway), Western/IP lysis buffer (Beyotime). The anti-MKK3 (Epitomics, #1728) antibody was used for IP.