Figure 6.

Hedgehog activation assay. The NIH-3T3 cells were transiently transfected with 1 μg sonic hedgehog (Shh) expression plasmid. For the inhibition of hedgehog signalling, cells were additionally exposed to 5-FU + BA combination and 10 μM cyclopamine (Cycl.) after 24 h transfection. Expression of the murine genes SHT, Gli1, Gli2, Ptch1, and Igf2 was determined after an incubation period of 48 h using quantitative real-time PCR in relation to the house-keeping gene TBP as a calibrator; *P < 0.05 (unpaired Student’s t-test).