Fig. 1

miR-184 induces OPC differentiation of NPCs. a The NPCs were cultured for 4 days in the oligo medium and then for 2 days in the growth factors free oligo medium. Top: NPCs were harvested and stained with antibodies against early- and late-stage OL markers, respectively. Scale bars, 50 μm. Bottom: Relative fluorescence intensity (G/B) representing the green fluorescence intensity (G) normalized to that of the blue one (B) was obtained after analysis of the images by ImageJ software. Unpaired t-test method was employed in each case to compare the amounts of results statistically. b Top: Expression of early- and late-stage OL markers analyzed by western blot. GAPDH was used as the control for normalization of protein bonds. Bottom: Average relative density of protein bands was obtained after densitometric analysis of the bands by ImageJ software followed by normalization to that of GAPDH as the internal loading control. One-way ANOVA method was used in each case to compare the amounts of results statistically. c miR-184 expression level evaluated by qRT-PCR in NPCs, 2 and 3 days after transduction with pLenti-III-Ctrl, pLenti-III-Scr, or pLenti-III-miR-184 relative to SNORD47 as an endogenous control. d, e qRT-PCR of lineage-specific genes from pLenti-III-Ctrl, pLenti-III-Scr, or pLenti-III-miR-184 transduced NPCs in the absence of growth factors at days 7 and 14. β-actin was used as internal control. f–h Relative expression levels of lineage-specific genes analyzed by qRT-PCR from pLenti-III-Ctrl, pLenti-III-Scr, or pLenti-III-miR-184-transduced NPCs at days 7, 14, and 21 in the presence of growth factors. β-actin was used as internal control. In the case of pLenti-III-Scr, transduction was performed with three pLenti-III-Scr construct and the values shown are the average obtained from them. Data represent mean ± SEM. Columns, mean of three replicates; bars, SD *P value < 0.05, **P value < 0.01, ***P value < 0.001. ns: non-significant (P value > 0.05)