Fig. 6

Knockdown of miR-200s impairs dopaminergic neurogenesis and radial migration in midbrain floor plate. a, b Immunofluorescence images show the distribution of GFP⁺ (green) cells at E13.5, which were electroporated at E11.5, with pCAG-Egfp control vector (a) or miR-200s sponge vector (pCAG-Egfp-Sponge; b) which antagonizes miR-200 family microRNAs. Boxes a, b in (a, b) are magnified in a and b, respectively, to show colocalization of GFP (green) and NR4A2 (magenta). White arrowheads indicated NR4A2⁺/GFP⁺ cells, while open arrowheads show NR4A2^-/GFP⁺ ones in (a, b. c) Percentage of GFP⁺ cells localized in MZ out of total GFP⁺ cells in floor plate of pCAG-Egfp (n = 5; median 61.4%, range 61.4–67.2%) or pCAG-Egfp-Sponge (n = 5; median 51.4%, range 44.4–55.0%) electroporated midbrain. d Percentage of NR4A2⁺/GFP⁺ cells out of GFP⁺ cells in floor plate of pCAG-Egfp (n = 5; median 47.0%, range 44.3–49.0%) or pCAG-Egfp-Sponge (n = 5; median 26.7%, range 21.1–33.7%) electroporated midbrain. e, f Immunofluorescence of TH (magenta) and GFP (green) in ventral midbrains electroporated with pCAG-Egfp control vector (e) or miR-200s sponge vector (pCAG-Egfp-Sponge; f). Boxes in e and f are magnified to show the colocalization of GFP (green) and TH (magenta). g Percentage of GFP⁺/TH⁺ cells in total GFP⁺ cells in floor plate of pCAG-Egfp (n = 5; median 10.1%, range 9.67–10.6%) or pCAG-Egfp-Sponge (n = 5; median 5.12%, range 4.41–5.81%) electroporated midbrain. Mann–Whitney U-test was used. Scale bar: 50 μm (b for a and b; e for e and f).