Distinct functions of HOXD proteins in regulation of neuronal differentiation.

<p>(<b>A</b>–<b>B</b>) Phase contrast imaging (<b>A</b>) and NEFM immunofluorescent staining (<b>B</b>) of BE(2)-C/Tet-Off/myc-HOXD cells cultured in the presence or absence of Doxy for 7 days. Scale bars, 50 µm. Shown are representative of three independent experiments with similar results. (<b>C</b>) Immunoblot analysis of PHOX2B and NEFM protein levels in the same cell samples described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040728#pone-0040728-g003" target="_blank">Figure 3</a>. Beta-actin levels are shown as loading control. Shown are representative of two independent experiments with similar results. (<b>D</b>) Quantification of <i>NEFM</i> mRNA levels in BE(2)-C/Tet-Off/myc-HOXD cells cultured in the presence or absence of Doxy for 3 days. The <i>NEFM</i> mRNA levels in BE(2)-C/Tet-Off/myc-HOXD cells in the presence of Doxy were designated as 1.0 (dashed line). The data were from two independent samples with each being assayed in triplicates and analyzed using two-tailed Student's <i>t</i>-test with the <i>p</i> values indicated. Error bars, SD.