Figure 9

In vitro assays for proliferation of GCPs from mutant cerebellum.

(A) GCPs from P7 cerebellum were cultured and immunostained using anti-NeuN and anti-BrdU antibody. More BrdU⁺ cells among the NeuN⁺ GCs were observed in the mutant GCPs compared with the wild-type GCPs. The rate of the BrdU⁺ cells among NeuN⁺ GCs significantly reduced after treatment with Shh inhibitor GDC-0049. (B) Quantitative analysis confirmed the increased BrdU⁺ cells among the NeuN⁺ GCs in the mutant GCPs (P = 3 × 10 ⁻⁶), whereas Shh inhibitor GDC-0049 reduced the BrdU⁺ cells among the NeuN⁺ GCs. These results indicated that loss of LKB1 increased the Shh-induced proliferation of GCPs (P = 4.2 × 10 ^− 3). Scale bars: 100 μm. The error bars indicate the SEM. *P < 0.05; **P < 0.01; ***P < 0.001 compared to the WT by Student’s t-test; n > 4 animals for each group.