Fig. 4

Hesperetin rejuvenates the naturally aged skin of WT mice. A For the hesperetin (Hes) treatment, 21-month old aged WT male mice were fed dietary hesperetin (100 mg/kg/day provided in food) or vehicle (Veh) control food (3.04% propylene glycol, w/w) for 5 months and then sacrificed at 26-months old. B Western blot analysis and quantification of Cisd2 protein levels in the skin of 26-months old mice treated with dietary hesperetin or vehicle control food for 5 months (starting at 21-months old). The protein level of Cisd2 in 3-months old WT mice serves as a young mouse control. C Masson’s trichrome staining of skin from the various different groups of mice. D Quantification of hair follicle density. The numbers of hair follicles present were normalized against the length measured. E Quantification of nuclear number of the sebaceous glands (SGs) in the skin of mice. F Quantification of the size of the sebaceous glands in the skin of mice. G Total reactive oxygen and nitrogen species (ROS & RNS) levels in skin tissue samples from young mice at 3-months old and Veh-treated or Hes-treated old mice at 26-months old. Data are presented as mean ± SD. *p < 0.05; **p < 0.005 by one-way ANOVA with Bonferroni multiple comparison test. UT untreated