Figure 4.
HuD antisense treatment (pDuH) blocks neurite outgrowth and GAP-43 expression in NGF-treated PC12 cells. Control and pDuH-transfected cells derived from the original PC12 cell line and the PC12-N21 clone were grown for 48 h in the presence or absence of NGF (100 ng/ml). Cells were then fixed with 4% PFA and processed for GAP-43 immunostaining using FITC-labeled secondary antibodies, as described in MATERIALS AND METHODS. (A) Phase contrast micrographs of untransfected PC12 cells and cells transfected with pDuH. Cells were photographed using an Axiovert Zeiss microscope at 400X magnification. (B) GAP-43 immunofluorescence in control and in pDuH transfected PC12-N21 cells. Images were taken using an Olympus BX-60 microscope at 400X magnification. Note that the HuD antisense RNA (pDuH) prevented neurite extension in both PC12 cell lines.