FIGURE 2.

Knockdown of LANP promotes neurite outgrowth in PC12 cells. PC12 cells were transfected with either control siRNA (A, C, F, and H) or siRNA targeting LANP (panels B, D, G, and I) both in the absence (A–E) and presence (F–J) of NGF. The cells were fixed and stained with anti-tubulin to monitor the contours of the cells to evaluate neurite outgrowth (compare panels A and B, F and G). Anti-LANP immunofluorescence demonstrates significant LANP depletion (compare panels C and D, H and I). These results are quantified (histograms E and J) by counting the number of cells displaying neurites longer than one cell body length. siRNA 1 and siRNA 3 represent two different siRNA duplexes targeting LANP. Scale bar = 20 μm. Cells were counted in 3 sets of high power fields representing triplicate transfections (typically 6 fields or more were counted with total number of cells >75 per set). Error bars represent S.E. ^*, p < 0.001. K, PC12 cells transfected with individual siRNA 1 and 3 show reduced protein expression by Western blot (WB) analysis. No reduction of LANP is found with control siRNA or cells transfected in a mock manner (no siRNA). A Western blot performed with an anti-actin antibody serves as a loading control.