Fig. 4: Notch1-induced IL-33 secretion increases whole-tissue ALDH1.
From: Endothelial Notch1 signaling in white adipose tissue promotes cancer cachexia
a, IPA comparative analysis of N1ICD-overexpressing AT-ECs and precachectic KPC AT-ECs identified IL-33 as a potential upstream regulator of transcriptomic changes. b, IL33 mRNA levels in N1ICD- compared to GFP-overexpressing human AT-ECs (n = 4–6 biologically independent experiments). c,d, Western blots (c) of human AT-EC IL-33 protein levels and quantification (d). Data were normalized to the expression of VCP (n = 6–8 biologically independent experiments). e,f, Analysis of Aldefluor activity in myeloid cells, including macrophages (CD45+CD11b+F4/80hi), monocytes (CD45+CD11b+Ly6G–Ly6C+), eosinophils (CD45+CD11b+SiglecF+) and neutrophils (CD45+CD11b+Ly6G+), by flow cytometry in vWAT (e) and sWAT (f) of NICDiOE-EC mice (n = 5–6 animals per group). Quantifications were normalized to each respective cell population. Flow cytometry experiments in e and f analyzing ALDHhi macrophages were performed twice with consistent results. Immunostainings and gatings for other ALDHhi myeloid cell populations were performed once. g, RT–qPCR analysis of Aldh1a2 mRNA expression in BMDMs treated with recombinant IL-33 for 72 h (n = 6 biologically independent experiments). h, Summary. WAT endothelial Notch1 mediates whole-tissue ALDH1 expression and RA production both directly and indirectly (via IL-33). Rald, Retinaldehyde. Data shown represent mean ± s.e.m. and were analyzed by unpaired, two-sided t-test with Welch correction.
