Fig. 4.

Atoh7 promotes the differentiation of retinal Müller cells into retinal ganglion cells. a Three days after induced differentiation, cells inside the neurospheres extended outwards and flattened; cell processes gradually grew in length; and the cells grew from the center to the periphery in a radial manner (×100); b Seven to ten days after induced differentiation, the cells reached the highest degree of differentiation; cells of varying sizes and shapes were observed. Some were flat and irregular with short processes, suggestive of glial cells; and the others had a small cell body and one or more long processes, resembling neurons (x 100). c Staining of DAPI, Thy1.1 and Brn3b of neurospheres; the right panel shows the merged images obtained for DAPI, Thy1.1 and Brn3b staining; d Bar chart shows the percentage of cells that were positive for both Thy1.1 and Brn3b retinal ganglion cell markers in the total differentiated cells. a–c indicates Atoh7 transfection, empty vector transfection and untransfection, respectively. e RT-PCR analysis of Thy1.1 and Brn3b expression in the total differentiated cells. a–c indicate Atoh7 transfection, empty vector transfection and untransfection, respectively