Extended Data Fig. 9: miR-126a-5p overexpression in vivo.
From: Muscle-derived miR-126 regulates TDP-43 axonal local synthesis and NMJ integrity in ALS models
a-b) Representative images of sciatic nerve cross section (a) and phrenic nerve in the diaphragm (b) of control animals injected with AAV PhP.Eb miR-126 viral particles. Green indicates GFP, red indicates NFH. Scale bars: 200 µm. c) Quantitative Taqman RT-PCR analysis of miR-126a-5p expression in SC, GC and brains of GFP-infected versus miR126-infected healthy mice. Data are shown as the mean relative miR-126a-5p level ± SEM. n = 3 mice in each condition. Two-tailed unpaired student’s t-test, *p = 0.0135(SC), 0.026(GC), 0.0363(Brain). d-e) Catwalk gait analysis of WT and SOD1G93A mice infected with either GFP or miR126, at symptomatic (p90) and late symptomatic (p130) stages. Data are shown as the mean stand duration (d; seconds) and % base of support on 4 limbs (e). n = 8 mice in each group. One-way ANOVA with Holm-Sidak correction. *p = 0.0277(d; p90; WT-GFP vs. SOD-GFP, and SOD1-GFP vs. SOD1-miR), p = 0.2587(d; p130; WT-GFP vs. SOD-GFP, and SOD1-GFP vs. SOD1-miR), **p = 0.0091(e; p90; WT-GFP vs. SOD-GFP, and SOD1-GFP vs. SOD1-miR), *p = 0.0143(e; p130; WT-GFP vs. SOD-GFP, and SOD1-GFP vs. SOD1-miR). f, g) Representative images (f) and quantification (g) of the hindlimb splay reflex in SOD1G93A mice and their littermates infected with either AAV PhP.Eb GFP, or with AAV PhP.Eb miR-126-GFP. Data are shown as the percent of mice who presented normal splay reflex ± SEM. n = 8; 8; 7; 7 mice in each group. Non-parametric Kruskal-Wallis test, *p < 0.02(WT-GFP vs. SOD1-GFP), *p = 0.0258(SOD1-GFP vs. SOD1-miR). h, i) In vivo longitudinal tracking of mouse weight (h) and survival (i) in all experimental groups. In (h), data are shown as the mean weight gain/loss at every timepoint compared with weight on injection day ± SEM. In (i), data are shown in a Kaplan-Maier survival curve starting at p150 (age of mice). n = (8,8;WT),(7,7;SOD1G93A) mice in each group. For a,b – images are representative for 3 similarly injected mice.
