Fig. 7.

Identification of miR10 targets in hNPCs. (A) Schematic overview of the experiment. Transduced hESCs were differentiated towards ventral MB for 14 days. The expression of miR-10 was induced by adding doxycycline to the media on days 11-14, whereas no doxycycline was added to the control groups. All groups include three replicates. (B) The fold change (FC) of each gene after miR-10a or miR-10b overexpression was plotted in a cumulative fraction graph. The cumulative distribution of the FC values for predicted miR-10 targets (red dotted lines; n=182) was significantly lower (shifted to the left) in comparison to that of all other genes, demonstrating that expression of miR-10 target genes is downregulated by expression of miR-10a/b (black lines; n=11,923, ***P<0.0001, Kolmogorov–Smirnov Z-test). (C) A large number of downregulated targets (89) were shared between miR-10a and miR-10b overexpression. Gene ontology (GO) analysis was performed on the shared targets, revealing miR-10 to be involved in functions related to transcription, actin cytoskeleton and ephrin receptor signalling. (D) The expression of the miR-10 target gene LIN28 was downregulated by overexpression of miR-10a and miR-10b. (E) Both WNT1 and MSX1 take part in the midbrain/hindbrain patterning and were downregulated upon overexpression of miR-10a and miR-10b.