Fig. 1: Age-associated meibomian gland atrophic changes in humans and mice.

a, Schematic of human eye emphasizing the tarsal plate. Infrared meibography images of upper and lower eyelids of young (29 years) and aged (62 years) men. yo, years old. Arrow indicates area of meibomian gland loss (right). b, Quantification of meibomian gland areas in upper and lower eyelids from nonpathological young (29–35 years; three men, three women) and aged (61–70 years; three men and three women) participants. ^*P = 0.0043 (top); ^#P = 0.0165 (bottom). AU, arbitrary unit. c, Outline of whole-mount meibomian gland staining of mouse eyelids. Pictures are representative of mouse upper and lower eyelids at 6 and 24 months of age. mo, months old. Arrow: area of meibomian gland loss. Scale bars, 1 mm. d, Quantification of meibomian gland areas of young (6 months, n = 12) and aged (24 months, n = 7) mice. ^*P = 0.0213. e, Local testosterone distribution in the meibomian gland using MS. Mouse eyelids were cut in a sagittal plane and counterstained with hematoxylin and eosin (H&E). Scale bar, 200 µm. Data are representative of four biologically independent eyelid samples with similar results. f, Outline of 3β-HSD activity measurements in the meibomian gland. Chromatograms show endogenous 3β-HSD activity arising from the meibomian gland of young (6 months) and aged (24 months) mice. 3β-HSD enzymatic activities were determined by measuring the conversion of ³H-DHEA (green) to ³H-androstenedione (magenta). c.p.m., counts per minute. g, Relative 3β-HSD activity in (f). Plots represent biologically independent samples of young (6 months, n = 8) and aged (24 months, n = 5) mice. ^**P = 0.0056. Data are the mean ± s.e.m. and were analyzed using unpaired two-sided Student’s t-test (b,d,g).