Research Paper Advance Articles

Figure 5. NDN knockdown in ex vivo iWAT abolishes the inhibitory effect of LKU4 on age-related adipose senescence. Eight-week-old C57BL/6J male mice were administered LKU4 or PBS daily for 4 weeks, followed by daily treatment of D-gal or 0.9% NaCl for an additional 8 weeks, with or without LKU4 supplementation. Mice treated with 0.9% NaCl, D-gal, and combined with LKU4 are designated as Con, D-gal, and D-gal/LKU4, respectively. Necdin was knocked down ex vivo in iWAT explants from D-gal/LKU4 mice using necdin-specific siRNA. (A) p53 acetylation in iWAT explants from each group of mice (n = 3). (B) SA-β-gal staining and protein levels of γH2AX in iWAT explants (n = 3). (C) Triglyceride (TG) levels in ex vivo iWAT explants from each group of mice (n = 3). (D) RT-qPCR analysis of senescence marker genes and lipid metabolism-related genes in iWAT explants (n = 3). (E) mtDNA copy number and CS activity (n = 3). (F) SASP mRNA levels in iWAT explants and SASP proteins secreted from iWAT explants (n = 3). The lowercase letters above the graphs indicate statistical significance at p < 0.05.