Immunocytochemistry analysis of SMAD6 in HepG2 cells. Samples were incubated with SMAD6 polyclonal antibody (Product # PA1-41026) using a dilution of 1:100 dilution overnight at 4°C followed by anti-rabbit DyLight 488 (Green) at a dilution of 1:500. Cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. Alpha tubulin was used as a co-stain at a 1:1,000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.