Supplementary Figure 7: Endogenous expression of synaptic target genes and Syt1 restoration (miR-137-Syt1-Venus).
From: The schizophrenia risk gene product miR-137 alters presynaptic plasticity
a–c. Endogenous target gene expression in the dentate gyrus and the mossy fiber-CA3 pathway of naïve C57BL/6 animals. a. Absolute endogenous mRNA levels measured by RT-qPCR. b–c. Endogenous protein levels of Cplx1, Nsf, Syn3 and Syt1 by b. western blot (cropped. Full-length blots are presented in Supplementary Fig. 9), and c. immunohistochemistry. Vibratome sections, immunolabeled against the mossy fiber pathway marker zinc transporter 3 (ZnT3, green), the target gene (magenta), and stained with nuclei dye DAPI (blue). Scale bar: 10 μm. The observation was repeated in at least three different animals. d. Additional ultrastructural images of the mossy fiber presynaptic terminal of ΔmiR-137OE-Venus (grey), miR-137OE-Venus (red), miR-137OE-Syt1-Venus (green), and ΔmiR-137OE-Syt1-Venus (purple) mice, black arrow: gold particles staining of Venus, red arrows: gap within the vesicle pool in miR-137OE-Venus, * active zone, scale bar: 100 nm. e. Total vesicle number for ΔmiR-137OE-Venus (grey, n=56), miR-137OE-Venus (red, n=53), miR-137OE-Syt1-Venus (green, n=56), and ΔmiR-Syt1-Venus (purple, n=58), H(3) = 77.75, ***p < 0.001. Multiple comparison two-tailed test after Kruskal-Wallis, *p < 0.05, box plot shows, in ascending order, the lowest maximum value, the first quartile, median, third quartile and the highest maximum value, dots are outlier. f. Nociception response measured by mean speed of movement during shock in (cm s–1), H(3) = 6.10, nsp = 0.1068. ns: not significant, s.e.m.
