Fig. 2: IF elicits AT stress response via adipocyte-autonomous p53 signalling. | Nature Communications

Fig. 2: IF elicits AT stress response via adipocyte-autonomous p53 signalling.

From: Adipocyte p53 coordinates the response to intermittent fasting by regulating adipose tissue immune cell landscape

Fig. 2

a Cell size quantification from histology images showing the normalised area of epididymal adipocytes of HFD-AL (n = 7 mice) and HFD-IF (n = 5 mice) groups. b, c mRNA expression levels of genes encoding for apoptotic, senescence, or DNA damage-associated markers in the adipocyte-rich fraction isolated from eWAT (b) or sWAT (c) of HFD-AL (n = 3 mice) or HFD-IF (n = 5 mice) mice. d Representative images showing histological stainings of cleaved caspase 3 in eWAT of HFD-AL and HFD-IF mice. Scale bars are 100 µm or 20 µm for the magnification. e Quantification of cleaved-caspase positive adipocytes in eWAT of HFD-AL (n = 10 mice) and HFD-IF (n = 7 mice) mice. f, g mRNA expression of Trp53 and p53 target genes Cdkn1a and Mdm2 in the adipocyte-rich fraction isolated from eWAT (f) or sWAT (g) of HFD-AL (n = 3 mice) and HFD-IF (n = 5 mice) mice. h Western blot analysis measuring p53 and GAPDH protein levels in isolated and differentiated cells from stromal vascular fraction (SVF) kept under nutrient-rich (Ctrl) or starvation (STV) conditions or treated with 1 µM of idasanutlin (NUT) (n = 3 independent isolations). i mRNA expression levels of Cdkn1a or Mdm2 in the differentiated SVF kept under Ctrl, STV or nutlin-treated conditions (n = 3 independent isolations). j Western blot analysis measuring p53 and GAPDH protein levels in differentiated p53 wildtype (WT) and knock out (KO) C3H10T1/2 cells (n = 3 independent experiments) kept under nutrient-rich control (Ctrl) or STV conditions. k, l mRNA expression levels of Cdkn1a and Mdm2 in p53 WT (k) and p53 knock out (KO, l) differentiated C3H10T1/2 cells kept under nutrient-rich control (Ctrl) or starvation (STV) conditions (n = 3 independent experiments). m, n mRNA expression levels of Trp53, Cdkn1a, and Mdm2 (m) or apoptotic genes (n) in differentiated SGBS cells under nutrient-rich control (Ctrl) or STV conditions and treated with siRNA targeting p53 (sip53) or siRNA control (siCtrl) (n = 3 independent experiments). Data are presented as mean values ± SEM. Significant differences were analysed by two-tailed, unpaired t-test (a–c, e–g), one-sample t-test (k, l), or one-way ANOVA (i, m, n) with Bonferroni post hoc tests. ***p < 0.001, **p < 0.01, and *p < 0.05. Exact p values: b Bax: 0.0004, Bcl2: 0.0086, Puma: 0.0235; c Bak1: 0.0194, Bax: <0.0001, Puma: <0.0001; f Trp53: 0.0364, Cdkn1a: 0.0013; g Cdkn1a: <0.0001, Mdm2: 0.0086; k Ckdn1a: 0.0111, Mdm2: 0.0070; m Ctrl siCtrl vs. STV siCtrl: Trp53: 0.0025, Cdkn1a: <0.0001, Mdm2: <0.0001; STV siCtrl vs. STV sip53: Trp53: <0.0001, Cdkn1a: <0.0001, Mdm2: 0.0021; n Ctrl siCtrl vs. STV siCtrl: Bcl2: 0.0353, Bax: 0.0013; STV siCtrl vs. STV sip53: Bcl2: <0.0001. Source data and uncropped blots are provided as a Source Data file.

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