Figure 5.

miR-342-5p Promoted Precocious Differentiation of NSCs In Vivo

(A) The forebrain areas of E15.5 mouse embryos were electroporated in utero with pcDNA6.2-GW/EmGFP-miR-342-5p (miR-342-5p) that could express miR-342-5p and EGFP, or with pcDNA6.2-GW/EGFP-negative control (Ctrl) that expressed only EGFP as a control. At E18.5, the mice were perfused and brains were fixed, cryosectioned, and stained with anti-PAX6 using immunofluorescence. Nuclei were counterstained with a Hoechst stain. Arrows and arrowheads represent EGFP- and PAX6-double-positive cells and EGFP-single-positive cells, respectively. Five pairs of littermates successfully electroporated with miR-342-5p and Ctrl separately were obtained.

(B) The percentage of PAX6-positive cells in EGFP-positive cells was compared (12 fields counted in each group).

(C) The EGFP-positive cells in each bin of five bins of the transfected cortical region were counted, and the percentage of EGFP-positive cells of each bin in total EGFP-positive cells was determined. Asterisks represent statistical analysis of the difference between the miR-342-5p-transfected and the control groups in the bin 1 and bin 2 areas (ten fields counted in each group).

(D) Samples in (A) were stained with anti-TBR2. The EGFP-positive cells were compared in the SVZ (TBR2+ zone), VZ (inside SVZ), and cortex (outside the SVZ) regions (ten fields counted in each group).

Bars, means ± SD. ^∗p < 0.05, ^∗∗p < 0.01, ns, not significant.