To determine whether PCAF regulates activation of the Hh pathway in HCC, we enhanced PCAF expression in the PLC/PRF/5 HCC cell line by transfecting a PCAF-expressing plasmid (Figure 1a) and decreased expression of PCAF in the Hep3B HCC cell line by transfecting siRNA targeting PCAF (Figure 1b).6 We measured the effect of PCAF on Hh pathway activity by measuring the expression of both GLI1 and Patched (PTCH1), which is used as a read-out for GLI activity, and also by measuring GLI-dependent reporter activity. Here, GLI1 is linked to hepatocellular carcinoma.