Figure 5: Hedgehog signaling inhibition attenuates carbachol-induced prostate cancer cell migration. Representative images of migrated PC-3 cells left (a) untreated or treated with (b) GANT61 at 3 μmol l-1, (c) carbachol at 10 μmol l-1, or (d) both carbachol and GANT61. Scale bars = 100 μm. (e) Quantification of migrated cells in the cell migration assays. Note that GANT61 (an inhibitor of hedgehog signaling) counteracted the carbachol-induced migration of PC-3 cells. Cells were counted in five random fields per section. Error bars indicate standard error. *P < 0.05; ***P < 0.001. (f) Proposed scheme for muscarinic receptor signaling transduction mechanisms in prostate cancer migration and invasion. Activation of CHRM1 through binding to Ach mediates PTCH1 and downstream GLI1 activation, resulting in gene transcription and ultimately promoting prostate cancer cell migration and invasion. Ach: acetylcholine; cAMP: cyclic adenosine monophosphate; PKA: protein kinase A; AKT: protein kinase B; GPCR: G protein-coupled receptors.