The Journal of Neuroscience, April 4, 2007, ():
Telomere Protection Mechanisms Change during Neurogenesis and Neuronal Maturation: Newly Generated Neurons Are Hypersensitive to Telomere and DNA Damage
J. Neurosci. Cheng et al.
27: 3722
Supplemental Data
Files in this Data Supplement:
- supplemental material
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Supplementary Figure 1. Characterization of neural cell cultures. A, Representative phase-contrast and confocal images of neuronal cultures established from E12 mice cerebral cortex at 2h, 2 days, 5 days after plating in differentiation medium. They display their different morphologies and the expression of molecular markers of cell identity. Nestin, a NPC marker and β3 tubulin (Tuj1), a neuronal marker. All cells were counterstained with propidium iodide (PI, red), a nuclear stain. B, The percentage of Nestin+ and Tuj1+ cells at different time point of plating were quantified and plotted. C, Western blot analysis of the molecular maturation of cortical neural cultures. Total protein extracts of E12 brain tissues, NPC cultures and neuronal cultures at different days in vitro (DIV) were analyzed using antibodies against NPC marker proteins (nestin and sox2) synaptic proteins (synapsin) and actin (to evaluate protein loading).
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Supplementary Figure 2. Target sequence 1 is the most effective in knocking down TRF2. A, four target SiRNA sequence and their location in mus musculus TRF2 mRNA (NM_009353). They are cloned in shRNA lentivirus plasmids (pLKO.1-puro) provided by sigma. B, 8DIV MN are infected with various lentivirus SiRNA for 48h and western blot analysis using anti TRF2 antibody. Lentivirus SiRNA with non-target sequence was used as control. Expression of β-actin was used as a loading control.
