Figure 2. TERT is co-localized with mitochondrial markers in the dendrites of cerebellar Purkinje neurons.

A. Representative image (n=4) of a cerebellar coronal section from an adult mouse immunostained with TERT (red) and Cytochrome C (green) antibodies. Arrows indicate examples of the co-localization of TERT and cytochrome C in the dendrites of Purkinje neurons (notice cell body in lower right corner). B. Representative image (n=7) of a cerebellar coronal section from an adult mouse immunostained with TERT (red) and COX-4 (green) antibodies. C. Example of a line profile analysis that plots the fluorescence intensity of both colors along the light blue line marked in B. Notice the co-variation of TERT (red) with the mitochondrial marker COX-4 (green). D. Representative image (n=3) of a cerebellar coronal section from an adult mouse immunostained with TERT (red) and parvalbumin (green) antibodies. Notice the colocalization of TERT and parvalbumin in the Purkinje neuron layer, but the lack of colocalization in the parvalbumin-positive cell bodies found within the molecular layer. E. Nuclear/cytoplasmic, mitochondrial and synaptosomal protein extracts were prepared from the cerebellum of eight mice (brains were pooled into groups of two in order to provide sufficient protein). Protein extracts (30 μg) were analyzed by immunoblot assay using antibodies against mTERT, synaptophysin and VDAC. The blot is a representative of 3 independent blots. F. The cerebellum was dissected from four 12–16 month old mice and protein extract were isolated from the following compartments: cytoplasm (Cyto), the nucleoplasm (NP, isolated nucleus treated with CHAPS alone) and protein bound to DNA (NDb, CHAPS with 1M NaCl). Protein extracts (30 μg) were analyzed by Western Blot assay using anti-TERT antibody, anti-Top1 antibody, anti-Lamin B and anti β-tubulin. Scale bar=20μm