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Figure 2. The lipid phosphatase null mutant PTEN impairs neuronal structure/neurite outgrowth. Rat cortical primary neurons were cultured for 2 wk and infected with sindbis viruses expressing GFP alone, GFP-IRES-wild-type PTEN or GFP-IRES-mutant PTEN. A) Expression of exogenous PTEN in infected primary neurons was confirmed by Western blots. Endogenous total tau and tubulin were also detected. B) Neurons expressing GFP alone (a–d), GFP-IRES-wild-type PTEN (e–h), or GFP-IRES-mutant PTEN (i–l) were visualized based on the GFP fluorescence (representing the expression of the transgenes), and further immunostained to detect tubulin (b, f, j) and tau (c, g, k). Fluorescence micrographs were visualized and recorded using a deconvolution microscope. d, h, and i are digitally merged images of tubulin and tau staining. C) Images of GFP-positive neurons were taken by fluorescence microscope (inset). The length and number of the neurons were counted. Results represent mean ± SE from at least 40 neurons of each experimental group. P > 0.05. Scale bars indicate 20 µm.
