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Fig. 3

miR-9 Potentiates Notch Activity to Maintain Quiescence

(A and B) Boxplots of qRT-PCR analysis showing the relative change in expression of the miR-9 transcripts and the Notch target her4 (A) and of the mature miR-9-5p/3p compared to a U6 small nucleolar RNA (snoRNA) control (B) after a 48 hr treatment with LY411575 or DMSO (control). The top and the bottom of each box indicate the 75th and 25th percentiles, respectively, and the dotted line indicates the median.

(C) Expression of miR-9 (magenta) in a GS+ (green)/PCNA+ (light blue) aNSC (yellow arrow) after a 48 hr LY treatment. Scale bar, 5 μm.

(D) Proportion of qNSCs (gfap+, MCM5; dark gray) and aNSCs (gfap+, MCM5+; white) in the miR-9+ or miR-9 NSC population after a 48 hr treatment with LY or DMSO.

(E) ISH for her4 2 dpi with the control (top) or miR-9 (bottom) vivoMO. Right panels are close-ups of the region within the red box. Scale bars, 100 μm (left); 50 μm (right).

(F) Epistasis experiment combining miR-9 kd with overexpression of Notch intracellular domain (NICD). The graph shows the percentage of qNSCs (dark gray) and aNSCs (white) in brains injected with the control or miR-9 vivoMO and in cells overexpressing NICD (NICD+) or not (NICD). p < 0.05, ∗∗p < 0.01; one-way ANOVA with Bonferroni post hoc correction. Data are represented as mean ± 95% CI; n = 3 brains per condition.

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Acknowledgments
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