Fig. 4: IL-33 expression is enhanced by endovascular injury-activated NFκB signaling pathway in perivascular MSCs. | Nature Communications

Fig. 4: IL-33 expression is enhanced by endovascular injury-activated NFκB signaling pathway in perivascular MSCs.

From: Perivascular mesenchymal cells instruct ST2+ reparative macrophages to promote endovascular injury-induced neointimal hyperplasia in mice

Fig. 4: IL-33 expression is enhanced by endovascular injury-activated NFκB signaling pathway in perivascular MSCs.The alternative text for this image may have been generated using AI.

a Volcano plot of differentially expressed genes (DEGs; p < 0.01, |Log2(Fold-change)|>1) FAI versus Sham based on bulk RNA-seq datasets. b UMAP plot showing Il33 expression in MSCs based on scRNA-seq datasets. c Representative images of the flow analysis of Il33-expressing cells in arteries of Il33GFP mice at D0 and D7. d Representative images of αSMA and IL-33 staining of arteries at D0 and D7. Scale bars, 50 μm. e mRNA levels of Il33 in the femoral arteries at D0 (n = 4), D2 (n = 8), D4 (n = 8), D7 (n = 8), and D14 (n = 4). f gMFI of GFP in MSCs using flow analysis (n = 6). g Plasma IL-33 levels in patients with or without in-stent restenosis (ISR, n = 25; Non-ISR, n = 25). h Schematic illustration showing the experimental design. HUVEC post-injured were incubated in fresh media for 24 h. CM were collected from HUVEC of normal (CM_Nor) or injured (CM_Inj). Boiled, CM_Inj post 95 °C incubation for 5 min; Frac<10K and Frac>10K, CM_Inj filtered into <10 and >10 kDa. i mRNA levels of Il33 in CM_Inj-exposed MSCs. j mRNA levels of Il33 in indicated CM-exposed MSCs for 24 h. k mRNA levels of Il33 in indicated CM-exposed MSCs for 24 hours. l Immunoblotting of IL-33 in indicated CM-exposed MSCs. m Signaling pathways enriched in MSCs of FAI group based on bulk RNA-seq datasets. n, Immunoblotting of indicated proteins in MSCs. o mRNA levels of Il33 in indicated CM-exposed in combination of JSH23 (100 μM) for 24 hours. p mRNA levels of Rela in MSCs transfected with siRela or siNC. q, mRNA levels of Il33 in indicated CM-exposed MSCs transfected with siRela or siNC. r Schematic illustration showing the 3 putative p65 protein binding sites (B.S.−1 ~ 3). ChIP-qPCR analysis showing the p65 binding to B.S.-1 ~ 3 in indicated CM-exposed MSCs for 24 h. Data are all shown as the mean ± s.e.m. *p < 0.01, **p < 0.01 or ***p < 0.001 by unpaired two-tailed Student’s t-test (g, h, p, q, r), or one-way ANOVA (e, f, i, j, k, o). Source data and statistic information are provided as Source data files.

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