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Figure 2 | Molecular Cancer

Figure 2

From: Paracrine sonic hedgehog signalling by prostate cancer cells induces osteoblast differentiation

Figure 2

LNShh cells upregulate Gli1 and Ptc1 expression in co-cultured MC3T3 cells. In this and subsequent graphs, gene expression in MC3T3 cells was determined by quantitative real time RT-PCR analysis using mouse species-specific primers shown in Table 1. (A) MC3T3 cells were mixed with either LNCaP or LNShh cells then seeded onto 6-well plates (mixed cultures). Expression of Gli1 and Ptc1 in MC3T3 cells cultured with LNShh cells (filled bars) relative to those cultured with control LNCaP cells (open bars) were compared. (B) MC3T3 cells were grown on 6-well plates in the presence of culture inserts containing either LNCaP or LNShh cells (separate co-cultures). Expression of Gli1 and Ptc1 in MC3T3 cells co-cultured with LNShh cells (hatched bars) relative to those co-cultured with control LNCaP cells (open bars) were compared. (C) Effect of cyclopamine inhibition of paracrine Shh signalling. MC3T3 cells were maintained for 7 days in mixed culture with either LNCaP or LNShh cells in serum-free culture media without or with 1 μM and 10 μM cyclopamine. Expression of Gli1 and Ptc1 in MC3T3 cells cultured with LNShh cells in the absence or presence of cyclopamine (filled bars) relative to those cultured with control LNCaP cells without cyclopamine (open bars) were compared. (D) Effect of exogenous Shh peptide. MC3T3 cells were cultured alone in serum-free culture media without or with 1 μg/ml Shh-N (modified active N-terminal peptide of human Shh; kindly provided by Curis, Inc.). Relative expression of Gli1 and Ptc1 in MC3T3 cells treated without (open bars) or with (filled bars) Shh-N were compared. Data are means ± SD of 2–4 assays. *, P < 0.05.

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