Figure 2
Recombinant human NRN1 increases neurite outgrowth in axotomized RGCs in vitro. Dissociated retinal cultures were treated with vehicle (control) or recombinant hNRN1 (200 ng/ml) for 10 days in culture. Fluorescence micrographs of neurite outgrowth in dissociated RGCs immunolabeled with Rbpms, Nefl and DAPI in (a) control and (b) NRN1-treated cells. Blue staining indicates DAPI labeled nuclei, green immunostaining for Rbpms and red for Nefl. Processed Rbpms-positive nuclear and Nefl neuronal tracings in (c) control and (d) NRN1-treated cells. Scale bar=50 μm. Photomicrographs were captured at 400 × original magnification. Quantification of average length of (e) neurites per RGC as described in a–d. Values represent the mean of total Rbpms-positive cells and total neurite length (μm) for four random images captured in 39 100 μm2 area for 15 wells from 4 independent experiments per group. Data are presented as mean±S.E.M. Statistical significance between treatment conditions determined by unpaired Student’s t-test, *P<0.05, n=15
