Research Paper Volume 11, Issue 15 pp 5628—5645

Rejuvenation of brain, liver and muscle by simultaneous pharmacological modulation of two signaling determinants, that change in opposite directions with age

Figure 4. Liver adiposity and fibrosis are reduced in old mice treated with Alk5i+OT. Livers were collected from non-injured mice and at 5 days post cardiotoxin-induced muscle injury, as illustrated in Figure 1C. (A) 10 µm liver sections were immunostained with Albumin+ (green) using Hoechst dye (blue) to label all nuclei, representative image is shown. (B) Albumin-negative fibrotic clusters (commonly found in old, but not young livers) were quantified; the incidence of fibrosis is reduced in the livers of old injured mice that were administered with Alk5i+OT, as compared to HBSS control **p=0.001, N for old control (Oc)=3, N for old+Alk5i+OT (OA5iOT)=3. (C) Isotype-matched IgG signal for albumin immunodetection was minimal. (D) Representative Oil Red O staining of liver sections show an age-specific increase in adiposity and reduction of old liver adiposity by the Alk5i+OT in both injured and uninjured animals. (E) Image J quantification of red pixel density in the Oil Red O assay was performed, as published (Rebo et al, 2016). Alk5i+OT diminished the liver adiposity of the old injured mice. N=6 in each injured cohort, ****p= 0.002 Oc & OA5iOT, N=6 in each uninjured cohort **p<0.001 Oc & OA5iOT. All scale bars=50 µm.