Extended Data Figure 9. 3DA treatment improves liver regeneration in aged mice.

a, Schematic representation of the experiment. Two-year old mice were treated 3 and 1 days before partial hepatectomy (PH) with 3DA or vehicle. The resected liver material was used for γH2AX staining and histopathology. 48h post PH, the rest of the liver was harvested and proliferation level was determined by Ki67 staining. b, Right side shows representative photographs of IF staining with antibody against ƴH2AX and fluorescent DNA stain (DAPI). The inlay shows a magnification of positive nuclei from the respective main photograph. Left side shows the quantification. A significantly higher amount (p<0.05) of γH2AX positive hepatocytes was detected in the control group (vehicle, n = 4) compared to experiment (3DA, n = 3), indicating a reduction in senescent cells. c, Right side shows representative photographs of IF staining with antibody against Ki67 and fluorescent DNA stain (DAPI). Left side shows the quantification. A significantly higher amount (p<0.05) of Ki67 positive hepatocytes were detected in experimental group (3DA, n = 3) compared to control (vehicle, n = 3), indicating that a reduction in senescent hepatocytes is associated with improved proliferation. Statistical significance was calculated using the unpaired two-tailed Student's t test. Error bars are represented as mean ± SEM; n represents number of mice. d-g, Pathological score (quantified blindly in a scale from 0-5) for the indicated parameters were assigned to H&E-stained liver sections from the experimental group (3DA, n = 4) and control group (vehicle, n = 4). Statistical significance was calculated using the unpaired two-tailed Student's t test. Error bars are represented as mean ± SEM; n represents number of mice. This figure was partly generated using Servier Medical Art, provided by Servier, licensed under a Creative Commons Attribution 3.0 unported license.