Figure 7

pX330-NeuN efficiently disrupts NeuN expression in vivo.

(a) Layer 2/3 neurons were co-transfected with 0.5 mg/ml pCAG-EGFP and 2.5 mg/ml pX330-NeuN using in utero electroporation at E15.5. Coronal sections were prepared at P4 and immunostained with anti-NeuN antibody (red). Arrows indicate normal NeuN expression in EGFP-positive neurons transfected with a pX330 control plasmid. Arrowheads indicate a dramatic reduction of NeuN expression in EGFP-positive neurons transfected with pX330-NeuN. Scale bar = 20 μm. (b) Histogram of the expression levels of NeuN in transfected neurons. The average NeuN signal intensity in EGFP-positive neurons transfected with a pX330 control vector was set as NeuN expression level 100. The number of neurons with NeuN expression level 100-110 and that of neurons with NeuN expression level <10 are also shown in (c,d), respectively. (c) Neurons with normal levels of NeuN expression were eliminated by pX330-NeuN. *p < 0.05; Mann-Whitney U-test (n = 4 pups for each condition). Error bars indicate SD. (d) The number of neurons with no NeuN expression was greatly increased by pX330-NeuN. **p < 0.01; Welch’s t-test (n = 4 pups for each condition). Error bars indicate SD.