Figure 4: CHRM1 stimulation promotes hedgehog signaling activation. (a) Western blotting images of GLI1, PTCH1, and SHH in PC-3 cells treated with 2 μg ml-1 carbachol for the indicated time. (b) Quantification of GLI1, PTCH1, and SHH expression in PC-3 cells treated with carbachol. (c) Images of GLI1, PTCH1, and SHH immunoblotting in PC-3 cells treated with 110 μg ml-1 pirenzepine for the indicated time. (d) Quantification of GLI1, PTCH1, and SHH in PC-3 cells treated with pirenzepine. Note that the expression of GLI1 and PTCH1 was mediated by CHRM1 activation, but that the expression of SHH was not significantly affected by carbachol or pirenzepine. (e) Real-time PCR results showed that GLI1 mRNA expression was suppressed in PC-3 cells treated with different concentrations of pirenzepine. (f) Quantitative analysis of PTCH1 mRNA expression in PC-3 cells after treatment with pirenzepine. (g) Quantitative analysis of SHH mRNA expression in PC-3 cells treated with pirenzepine. Each sample was run in triplicate. The results are shown as the mean ± s.e.m. GLI1: GLI family zinc finger 1; PTCH1: patched 1; SHH: sonic hedgehog; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; s.e.m.: standard error of the mean.