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. 2014 Jun 26;7:59. doi: 10.3389/fnmol.2014.00059

FIGURE 1.

FIGURE 1

Either a constitutively active Pea3-VP16 fusion or Pea3 induced with EGF can induce neurite formation in PC12 cells. (A) A VP16 fusion of mouse Pea3 leads to neurite formation in PC12 cells. Cells were transfected with 500–1 μg of GFP expressing plasmid in 1:3 ratio with either a control plasmid expressing VP16 acidic activation domain alone, or a fusion of Pea3 to VP16 activation domain, and analyzed by fluorescence microscopy. A typical micrograph is shown. (B) The cells were scored for % neurite formation among the GFP-expressing population. (Two different DNA preparations were used for Pea3VP16 plasmid; data shown for only one of these.) (C) Luciferase assays in response to NGF, EGF, and transfected Pea3 on SRE-Luciferase reporter construct; average of at least three independent experiments. (D) A typical micrograph of cells transfected with 500–1 μg of GFP expressing plasmid in 1:3 ratio with either a control plasmid expressing pCDNA3 (not shown), pCDNA3 Pea3, or pCDNA3 Pea3 in combination with EGF as described, and analyzed by fluorescence microscopy. The arrow shows axon-like processes several cell body length. (E) A graph summarizing the scoring for differentiation experiment in (D). The experiment was repeated with two different DNA preparations for pCDNA3 Pea3, and at least 3 fields per well were counted. The control plasmid pCDNA3 yielded no differentiation (not shown), while NGF in a similar time scale (2–4 days after transfection) resulted in around 12% differentiation (not shown).