Extended Data Fig. 4: UVB radiation induces ghrelin production and secretion in skin adipocytes, Related to Fig. 3.
From: Food-seeking behavior is triggered by skin ultraviolet exposure in males
a, Plasma protein levels of insulin upon UVB (50 mJ/cm2) or mock-UVB (control) -irradiated mice at 10 weeks after the first treatment (Biologically independent males: n = 10 control and n = 12 UV biologically independent mice per condition; Biologically independent females: n = 10 control and n = 9 UV mice per condition). b, Stomach tissue from control, UVB-irradiated (50 mJ/cm2), and 22-h-food-deprived mice after 5 weeks of treatment. Immunofluorescence for ghrelin (red), perilipin 1 (Plin1, an adipocyte marker, green), and nuclei (DAPI, blue). Ghrelin intensity normalized to DAPI (n = 4 biologically independent mice per condition). c, Relative and arbitrary mRNA levels of pcsk1, GOAT, and ghrelin from indicated tissue from UVB (50 mJ/cm2) or mock-UVB (control) irradiated male mice at 10 weeks after first treatment and from LiSa-2 differentiated adipocytes 24 h after UVB (50 mJ/cm2) radiation. Data normalized to 36b4 (n = 4 biological replicates). d, Relative p53 and p21 mRNA levels from differentiated 3T3-L1 adipocytes 24 h after UVB (50 mJ/cm2) or control radiation. Data normalized to 36b4. e, Representative 3D construction of immunofluorescence image of human skin adipose tissue of UVB (2,000 mJ/cm2) 24 h post-treatment stained for ghrelin (red), perilipin 1 (Plin1, an adipocyte marker, green), and nuclei (DAPI, blue). f, Skin tissue from (b). Upper panel: Representative images of immunofluorescence. Bottom panel: Ghrelin intensity normalized to DAPI (n = 3 biologically independent mice per condition). g, Left panel: Immunofluorescence of differentiated 3T3-L1 adipocytes after 24 h of UVB (50 mJ/cm2) or control treatments stained for ghrelin (red) and nuclei (with DAPI, blue). Right panel: Ghrelin intensity normalized to DAPI intensity (n = 9 random fields from 3 biological replicates). h, Arbitrary of Agrp and Npy mRNA levels from brain hypothalamus upon 10 weeks of mock-UVB (control) treatment (n = 3 biologically independent mice per condition). Data normalized to 36b4. In all relevant panels: Data are presented as mean ± SEM; Two-tailed unpaired t-test p-values are shown, or statistical details for sex or UVB or treatment factors in the ANOVAs (F-values, degrees of freedom, p-value) with interaction appears in Supplementary Table 10, or two-way ANOVA analysis with multiple correction test appears in Supplementary Table 11.
