Requirement of Dpy19L1 in neurite extension of cortical neurons.

<p>(A) COS-7 cells were transfected with either a Dpy19L1 siRNA or control siRNA along with a CAG-Dpy19L1 expression plasmid, followed by western blot analysis at 48 h after transfection. Dpy19L1 expression was efficiently reduced by Dpy19L1 downregulation. α-Tubulin was used as a control. (B) Either control siRNA or one of two Dpy19L1 siRNAs was transfected in embryonic cortical neurons, and 48 h later, the expression level was checked by RT-PCR. Each Dpy19L1 siRNA decreased <i>Dpy19L1</i> expression. β-actin was used as a control. (A, B) The blots (A) and the gels (B) were cropped from the same gels, and the full-length blots and gels are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0167985#pone.0167985.s005" target="_blank">S5B and S5A Fig</a>, respectively. (C−E) E14.5 cortical neurons were transfected with control or Dpy19L1 siRNA and allowed to differentiate for 72 h. The lengths of the longest neurite were compared between control and Dpy19L1-downregulated neurons. Dpy19L1-downregulated neurons show reduced neurite length compared with that of control neurons. Scale bar: 100 μm. (D) Histogram shows distribution of neurite lengths of control and Dpy19L1-downregulated neurons. (E) Average neurite length of control siRNA and Dpy19L1 siRNA transfected neurons. (F) Comparison of the number of cleaved Caspase3-labeled cells between control and Dpy19L1-downregulated neurons at 48 h after transfection. All data shown here are from at least three independent culture experiments. Values are mean ± SEM. *<i>P</i> < 0.05.