Anti md2

HCC cells were lysed by RIPA buffer (Thermo Fisher Scientific, USA). Protein concentrations were determined by Bradford protein assay. Equal amounts of proteins were separated on SDS‐PAGE (CA) followed by electro‐transferring on the PVDF membrane (Bio‐Rad, CA). After finishing the electro‐transfer, membranes were immersed in the blocking buffer containing 5% BSA for 2 h prior to the incubation with primary antibodies, including anti‐TLR4 (Cat: MAB6248, R&D Systems, USA), anti‐MD2 (Cat: ab24182, Abcam, USA), anti‐glypican‐3 (Cat: ab66596, Abcam, USA), anti‐Sp1 (Cat: 9389, CST, USA), anti‐STAT3 (Cat: 9139, CST, USA), anti‐phospho‐STAT3 (Cat: 9145, CST, USA), anti‐HIF‐1α (Cat: 36169, CST, USA), anti‐phospho‐p38 MAPK (Cat: 4511, CST, USA), anti‐IκB‐α (Cat: 4812, CST, USA), anti‐p65 (Cat: 8242, CST, USA) and anti‐MyD88 (Cat: 50010, CST, USA), respectively. HRP‐conjugated secondary antibodies, including anti‐rabbit IgG (7074, CST, USA) and anti‐mouse IgG (7076, CST, USA), were correspondingly used to the incubate the membranes for 2 h (Cell Signaling Technology, USA). Bands were visualized under the chemiluminescence system (Bio‐Rad, USA).