Fig 1: MSCs release some cytokines in vitro such as IL1β, IL6, and IL8 in different inflammatory scenarios and their migration potential is maintained only until P2. MSCs were treated for 24 h as indicated and the presence of IL1β, IL6, and IL8 were measured in their supernatants (A–C). The mRNA levels of MMP2 and MMP9 (D) and of TIMP1 and TIMP2 (E) were analyzed after treatments to compare from P1 to P4. The relative activity of MMP2 was analyzed by zymography in gel (F) in control and LPS+IFN treated cells from P1 to P4. The migration capacity of cells was measured using a “wound healing” assay (G) at 0, 24 h and 7 days in control cells at P1 and P4. *p < 0.05; **p ≤ 0.01 vs. the control or P1 condition.
Fig 2: TAS1R3 promotes intestinal inflammation in patients with IBD. a Transcriptome meta-analysis. Only datasets preprocessed and normalized in one step were used (GSE160804/GSE126124/GSE95095/GSE75214/GSE53306). b PCA of intestinal biopsy transcriptomes from patients with and without IBD. Grey and red points represent individual IBD (n = 204) and non-IBD (n = 74) samples, respectively. c Hierarchical clustering and heatmap of up- and downregulated genes. A total of 1,697 significantly DEGs were identified (unpaired Student’s t-test; FDR P < 0.001). d TAS1R3, e MTOR, and f PPARG mRNA expression in intestinal biopsies from patients with (n = 204) and without IBD (n = 74). Spearman’s correlation analysis of associations between intestinal TAS1R3 mRNA expression and that of g MTOR and h PPARG, as well as pro-inflammatory cytokines: i IL1B, j TNFa, and k IL8. All correlations shown are significant. Data are expressed as means ± standard errors of the mean. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 (unpaired Student’s t-test). DEF, differentially expressed gene; FDR, false discovery rate; IBD, inflammatory bowel disease; PCA, principal coordinate analysis
Fig 3: TAS1R3 controls pro-inflammatory cytokine production and secretion by regulating the mTOR–PPARγ axis. Western blotting of (a) phospho-mTOR, mTOR, and PPARγ expression in ND- and WD-fed Tas1r3−/− and Tas1r3+/+ mice and (b) and (c) densitometric data (n = 6 mice/group). Representative images of at least three different blots and α-tubulin. d–f NCI-H716 cells were transfected with TAS1R3 (10 nM) or scrambled control (10 nM) siRNA for 48 h and stimulated with or without fructose (10 mM), glucose (10 mM), and palmitate (10 μM) for 12 h. qRT-PCR analysis of (d) MTOR and (e) PPARG mRNA expression relative to GAPDH (n = 5/group). g–i NCI-H716 cells were pretreated with the TAS1R3 antagonist, lactisole (2.5 mM), for 30 min and stimulated with fructose (10 mM), glucose (10 mM), and palmitate (10 μM) for 12 h. qRT-PCR analysis of (g) MTOR and (h) PPARG mRNA expression relative to GAPDH (n = 5/group). f and i PPARγ protein abundance was assessed by western blotting (n = 6/group). Alpha-tubulin was used as the loading control. j–m NCI-H716 cells were transfected with TAS1R3 (10 nM) or scrambled control (10 nM) siRNA for 48 h and stimulated with the PPARγ antagonist, GW9662 (10 μM), in the presence of fructose (10 mM), glucose (10 mM) and palmitate (10 μM) for 12 h. qRT-PCR analysis of (j) TNFA and (k) IL8 mRNA expression relative to GAPDH (n = 5/group) and enzyme-linked immunosorbent assay of (l) TNF-α and (m) IL-8 protein abundance (n = 3/group). Data represent means ± standard errors of the mean of three to five independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 (one-way analysis of variance followed by Bonferroni post-hoc test). ND, normal diet; qRT-PCR, quantitative reverse transcription PCR; TAS1R3, taste receptor type 1 member 3; WD, Western diet
Supplier Page from Abcam for Human IL-8 ELISA Kit