Journal: Disease Models & Mechanisms

Article Title: Bmi1 overexpression in the cerebellar granule cell lineage of mice affects cell proliferation and survival without initiating medulloblastoma formation

doi: 10.1242/dmm.009506

Figure Lengend Snippet: Cell cycle length is increased in Math1-Bmi1 GCPs. (A,B) Sagittal sections at P15 of Math1-Bmi1× Math1-EGFP cerebella (TG) and Math1-EGFP littermate control (WT), showing the presence of an EGL and Math1-GFP-positive cells in both genotypes. (C,D) Sagittal sections of EGL showing EdU/BrdU double labelling at P8 in wild-type (C) and Math1-Bmi1 (D) cerebella. Arrows show examples of double-positive cells. (E,F) Images in C and D with DAPI overlay for labelling of nuclei. Scale bars: 0.1 mm. (G) Bar chart showing a decrease in the mean EdU/BrdU double-positive index (double-positive cells per μm 2 of EGL) ± 1 s.e.m. in transgenic animals compared with wild-type littermates. (H,I) Quantification of gene expression changes by qRT-PCR array at P6, expressed as mean fold change in comparison with wild type ± 1 s.d., showing a trend of downregulation in positive regulators of the cell cycle (H) and DDR pathway genes (I). Asterisks indicate significant downregulation compared with wild type. EGL, external granule cell layer; IGL, internal granule cell layer; TG, transgenic; WT, wild type; PI, propidium iodide.

Article Snippet: Sections were mounted with either VECTASHIELD mounting medium containing DAPI (Vectorlabs Inc.) or treated with 5 μg/ml propidium iodide (Sigma, UK) and 10 μg/ml RNase A (Sigma, UK) for 20 minutes prior to mounting with CITIFLUOR™ (Citifluor Ltd).

Techniques: Transgenic Assay, Expressing, Quantitative RT-PCR