Figure 3.

miR-218 promotes mESC differentiation into DAn. a, Schematic representation of the experimental procedure of mESCs infected with a DOX-inducible LV expressing miR-218-1 upstream of an Ires-GFP sequence (or Ires-GFP empty vector), FACS-purified and amplified, before DA differentiation. b, TaqMan assay for miR-218 from FACS-purified mESCs infected with LVs for miR-218-Ires-GFP or empty Ires-GFP. Data were normalized to the average of the reference sno-202. Data are mean ± SEM of 2^−ΔCt values. *p < 0.05, n = 3 (Student's t test). c, The expression of dopaminergic genes (Th, Pitx3, Nurr1, and Lmx1a) was analyzed by qPCR, during the differentiation protocol in the presence or absence of DOX. Data are mean ± SEM from three independent experiments. *p < 0.01 (Student's t test). d, Immunostaining for Th in FACS-sorted, amplified, and differentiated in mESCs infected with LVs for miR-218-Ires-GFP or an empty Ires-GFP, as control, in presence of DOX. A panel of 32 low-magnification images (5×) for each condition is shown.