Figure 3.

Validation of Lentiviruses to Overexpress/Downregulate miR-135a in NPCs In Vitro and In Vivo

(A) Relative expression levels of mature miR-135a in primary NPCs transduced in vitro with lentivirus transcribing the immature short-hairpin precursor of miR-135a (sh-miR-135a, i.e., gain of function), or a sponge for miR-135a (sponge miR-135a, i.e., loss of function), or control viruses expressing scrambled RNAs.

(B) Representative micrographs showing immunostaining for Nestin-CFPnuc (green), BrdU-positive cells (red), and GFAP-positive cells (white), and nuclear DNA with DAPI (blue) in the hippocampal SGZ of 6- to 8-week-old Nestin-CFPnuc mice, injected with lentiviruses (same used in A), kept 10 days under standard conditions and subjected to three injections of BrdU 24 h before sacrifice.

(C) Representative micrographs showing immunostaining for Nestin-CFPnuc (green), BrdU-positive cells (red) and nuclear DNA with DAPI (blue) in the hippocampal SGZ of 6- to 8-week-old Nestin-CFPnuc mice, injected with miR-135a sponge or scrambled-sponge lentivirus, kept 10 days under standard conditions, and subjected to three injections of BrdU 24 h before sacrifice.

(D) Percentage of BrdU and Nestin-CFPnuc double-positive cells over total BrdU⁺ cells in the SGZ of mice injected with lentiviruses.

Data are expressed as means ± SEM, n = 7 mice per group. One-way ANOVA Bonferroni as post hoc. ^∗∗p < 0.01, ^∗∗∗p < 0.001. Scale bars, 50 μm (B) and 25 μm (C).