Immunocytochemical staining procedures were described previously38 (link). Briefly, 1 × 105 HepG2 cells were seeded on a glass slide overnight and rinsed once with PBS. −20 °C methanol was added immediately for fixation and washed with PBS. Detection of target proteins was performed according to the manufacturer’s instructions. Primary specific Anti-p-STAT3 antibody was purchased from Santa Cruz, CA, USA and specific antibodies to Ki67, PCNA, PSA, IL-6, IL-6R, gp130, EGFR and TGFβ were gifts from Maixin biotech., Fuzhou, China.
Paraffin sections were stained by using standard immunohistochemical procedures described previously. Briefly, mouse tissue samples were fixed with 10% formalin for 24 hrs and paraffin embedded. Four mm thick serial sections were mounted on glass slides and were immunostained with various antibodies described in the previous section following standard procedures39 (link).
Paraffin sections were stained by using standard immunohistochemical procedures described previously. Briefly, mouse tissue samples were fixed with 10% formalin for 24 hrs and paraffin embedded. Four mm thick serial sections were mounted on glass slides and were immunostained with various antibodies described in the previous section following standard procedures39 (link).