Fig. 6: Estrogen blocks p53 transcriptional activation of ghrelin upon UVB exposure.
From: Food-seeking behavior is triggered by skin ultraviolet exposure in males
a, Estrogen levels in human skin adipose tissue (n = 5 and n = 7 independent human male and female donors, respectively). b, Left: Experimental design. Right: Representative images of cells stained for ghrelin (red), perilipin 1 (Plin1, an adipocyte marker, green) and nuclei (with DAPI, blue) (n = 10 random fields from 3 biologically independent samples per condition). Relative ghrelin intensity was normalized to DAPI. c, ER-α protein levels in differentiated LiSa-2 adipocytes 24 h after UVB (50 mJ/cm2) or mock-UVB (control) irradiation treatments. β-actin was used as the loading control. Quantification of the protein amount normalized to β-actin (Q). d, mRNA levels of p53, p21 and ghrelin in differentiated LiSa-2 adipocytes 24 h post treatment with 100 nM DHT or 100 nM β-E2 or vehicle and UVB (50 mJ/cm2) or mock-UVB (control) irradiation (n = 3 biologically independent samples per condition). Data normalized to 36b4. e, Schematic representation of ghrelin promoter with p53 binding regions. f,g, p53 (f) and NCOR1 (g) occupancy over human ghrelin promoter in differentiated LiSa-2 adipocytes treated as in (c) (n = 3 biologically independent samples per condition). ChIP levels (fold) normalized to input. h, Experimental design. i, PhenoTyper analysis of weekly food intake (in grams) of indicated female mice as in (h) (Week 1 and 2: n = 12 biologically independent mice per condition; Week 3: n = 12 biologically independent mice per sham condition, n = 10 biologically independent mice for OVX control and n = 11 biologically independent mice for OVX UV; Week 4: n = 11 biologically independent mice per condition). j, Staircase test for indicated female mice and treatments as in (h) (n = 12 biologically independent mice per condition). k, Open-field analysis by the indicated mice and treatments (h) (n = biologically independent 12 mice per condition). Right panel: Representative heat maps. l, Relative ghrelin mRNA levels in OVX or sham female mice skin, after 5 weeks of daily UVB (50 mJ/cm2) or mock-UVB (control) exposures (n = 5 biologically independent mice per condition). Data normalized to 36b4. m, Upper panel: Representative immunofluorescence images stained as in (b), in skin tissue from (l). Ghrelin intensity normalized to DAPI (n = 10 images from 3 biologically independent mice per condition). n, Relative ghrelin mRNA levels in human female skin adipose tissue 25 h post treatment with estrogen inhibitor (leterozole (5 µM)) or vehicle (DMSO) and a single UVB (500 mJ/cm2) or mock-UVB (control) irradiation session (n = 4 biologically independent human donors per condition). Data normalized to 36b4. o, Relative ghrelin mRNA levels in differentiated primary human female adipocytes after treatment as in (n) (n = 5 biologically independent samples 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 OVX or UVB 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.
