Fig. 4

Flow cytometry-sorted neurons show normal morphologies in culture. (A) Tg(elavl3:eGFP)knu3 embryo (lateral view, 2 dpf). Scale bar, 50 µm. (B) Spinal cord of a Tg(elavl3:eGFP)knu3 embryo (dorsal view, 2 dpf). Scale bar, 50 µm. Anterior is always to the left. (C) Dissociated primary cells of 2 dpf wild type (n = 1 preparation) and elavl3:eGFP embryos (n = 3 preparations) gated by the settings described in Fig. S2 were further gated and sorted regarding GFP-expression and plotted as histogram. Virtually no GFP-positive (GFP+) cells were detected in the wild type preparation, meaning that all GFP+ cells of the elavl3:eGFP preparations indeed expressed the transgene. The amount of GFP+ cells was comparable in all three elavl3:eGFP preparations. (D)elavl3:eGFP-positive cells prepared with the standard protocol for primary cell culture (1 dap). Scale bar, 10 µm. (E) Same morphologies as observable in (D) can be found among elavl3:eGFP-positive cells which have been selected by flow cytometry and cultured for 1 day. Scale bar, 10 µm. Shown are collections of the images of (D) two and (E) three preparations with comparable results.