Figure 4.

miR-342-5p Promoted the Differentiation of NS/PCs into INPs In Vitro

(A) NS/PCs were transfected with miR-342-5p mimics or control (five independent transfections performed), and cultured under the adherent condition to proliferate at clonal density for 7 days. The newly formed colonies were further induced to differentiate for 48 hr with low mitogens (DF12 containing 1/2 × N², 5 ng/mL bFGF and 5 ng/mL EGF) supplemented with 5% FBS. Cells were then fixed and stained with anti-GFAP and anti-TUJ1. The frequencies of GFAP+, TUJ1+, and GFAP+ TUJ1+ colonies were compared (50 colonies counted in each group).

(B and C) Adherently cultured NS/PCs were stained with anti-PAX6 and anti-NESTIN (B), and percentages of PAX6+ cells in total cells or NESTIN+ cells were determined (C) (eight fields counted in each group).

(D and E) NS/PCs were stained with anti-TBR2 and anti-NESTIN (D), and percentage of TBR2+ cells were determined (E) (eight fields counted in each group).

Bars, means ± SD. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.