Extended Data Fig. 9: Observed rejuvenation effects of global aging miRNAs and validation experiments for predicted targets of miR-29c-3p.

z-scored expression in the healthy aging cohort (TMS) for every timepoint and z-scored expression in the parabiosis cohort for the four different groups (IY, HY, HA and IA) for miR-184-3p in the liver (A) and miR-300-5p in GAT (B). The first to the third quartile are covered within the boxes with the median value shown as line inside the box. Maximum and minimum values are shown as whiskers or values up to 1.5-times the interquartile range above and below the first or third quartile if outliers are present. (C) Measured RLU (relative light unit) in percent for all target constructs for miR-29; reduction to under 80% determines a high confidence target, reduction between 80-90% low confidence target (n = 4 biologically independent experiments for each target, each as technical duplicates). Color coded in dark green for significant reduction with a p-value ≤ 0.01 (ADAMTS17_1: 0,0000004, ADAMTS17_2: 0,00000007, ADAM12_1: 0,00001, LOX_2: 0,0006, LOX_3: 0,000000001, VASH1_2: 0,0067), light green for significant reduction with a p-value ≤ 0.05 (AD-AM12_2: 0,0306) and in light brown for non-significant reduction with a p-value ≥ 0.05 (n.s; APLNR: 0,1485, LOX_1: 0,1892, VASH1_1: 0,5457). P-values were calculated by an unpaired t-test. Data are shown as mean ± SD. (D) Control experiment results luciferase assay, negative control: vector alone and vector with miRNA and positive control with vector (pMIR+ pSG5; pMIR+miR-29a, PC miR-29a-3p + pSG5) and positive control (PC miR-29a-3p + miR-29a) (n = 4 biologically independent experiments, each as technical duplicates; *** = p-value ≤ 0.001, n.s. = p-value ≥ 0.05). P-values were calculated by a Welsh’s t-test (pMIR+pSG5 vs. pMIR+miR-29a = 0,6314; PC miR-29a-3p+miR-29a vs. PC miR-29a-3p + pSG5 p-value < 0,0001; PC miR-29a-3p+miR-29a vs. pMIR+miR-29a p-value < 0,0001.) Data are shown as mean ± SEM.