Figure 3

The sonic hedgehog signaling pathway was regulated by lymphangiogenic factors. (A) H1299 and Calu-1 cells were treated with 500 ng/ml N-Shh for the indicated times. The total RNA was harvested and subjected to RT-PCR using lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1), vascular endothelial growth factor D (VEGF-D) or β-actin (loading control) primers. (B) H1299 cells were treated with 500 ng of N-Shh or 10 μM cyclopamine-KAAD for 24 hours, and RNAs were isolated and subjected to RT-PCR for Glioma-associated oncogene homolog zinc finger protein 1 (Gli1), LYVE-1, VEGF-D or β-actin (loading control).