Fig. 3: Notch1 regulates RA metabolism through ALDH1 expression.
From: Endothelial Notch1 signaling in white adipose tissue promotes cancer cachexia
a, Overlapping predicted upstream regulators of transcriptomic changes in KPC (precachectic) and N1ICD-overexpressing AT-EC datasets compared to their respective controls. b, GO term ‘cellular response to RA’ enrichment plot of cachectic versus healthy vWAT (GSE131835). c,d, Intracellular ATRA levels in human vWAT ECs (n = 14 biologically independent experiments; c) and sWAT ECs (n = 12 biologically independent experiments; d) treated with AdN1ICD or AdGFP were measured by mass spectrometry. e, vWAT ECs overexpressing AdN1ICD or AdGFP were analyzed by ChIP–seq using an antibody to H3K27ac. N1CD increased H3K27ac at the HEY2 locus (left, red box) and at the ALDH1A2 locus (right, red box). RBP-J binding motifs are identified within the regions associated with increased H3K27ac after N1ICD overexpression at both the HEY2 and ALDH1A2 loci. RBP-J binding motifs are highlighted by the gray boxes; Mb, megabases. f,g, mRNA levels of ALDH1 isozymes (n = 3–4 biologically independent experiments; f) and ALDH1A2 protein levels (g) analyzed by western blotting in human AT-ECs overexpressing AdN1ICD or AdGFP; H-vWAT, human vWAT; H-sWAT, human sWAT. h, Western blots were quantified and normalized to VCP (n = 4 biologically independent experiments). i, mRNA expression of Aldh1 isozymes in NICDiOE-EC AT-ECs (n = 3 biologically independent experiments). Data shown represent mean ± s.e.m. and were analyzed by Wilcoxon test (c and d) or unpaired, two-sided t-test with Welch correction (f, h and i).
