SUCNR1 mediates inflammasome priming in intestinal epithelial cells. (a) HT29 cells treated with succinate 1mM and/or an inflammatory cocktail. Graphs show mRNA (n = 7) and protein (n = 3) expression of SUCNR1. Image of a representative Western Blot of one independent experiment. Bars in graphs represent mean ± SEM; * p < 0.05 and ** p < 0.01 vs. vehicle cells. (b-e) HT29 cells transiently transfected with a specific siRNA against SUCNR1 or ctrl; (b) Graphs show mRNA and protein expression of SUCNR1 (n = 5). (c) Graphs show mRNA expression of NLRP3, CASP1, IL1B, ASC and IL18 (n = 5). (d) Graph shows protein expression of pro-Caspase-1 (n = 4). Image of a representative Western Blot of one independent experiment. (e) Graph shows secreted protein levels

SUCNR1 mediates inflammasome priming in intestinal epithelial cells. (a) HT29 cells treated with succinate 1mM and/or an inflammatory cocktail. Graphs show mRNA (n = 7) and protein (n = 3) expression of SUCNR1. Image of a representative Western Blot of one independent experiment. Bars in graphs represent mean ± SEM; * p < 0.05 and ** p < 0.01 vs. vehicle cells. (b-e) HT29 cells transiently transfected with a specific siRNA against SUCNR1 or ctrl; (b) Graphs show mRNA and protein expression of SUCNR1 (n = 5). (c) Graphs show mRNA expression of NLRP3, CASP1, IL1B, ASC and IL18 (n = 5). (d) Graph shows protein expression of pro-Caspase-1 (n = 4). Image of a representative Western Blot of one independent experiment. (e) Graph shows secreted protein levels

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Intestinal epithelial cells (IECs) constitute a defensive physical barrier in mucosal tissues and their disruption is involved in the etiopathogenesis of several inflammatory pathologies, such as Ulcerative Colitis (UC). Recently, the succinate receptor SUCNR1 was associated with the activation of inflammatory pathways in several cell types, but li...

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Context 1
... treatment with succinate-as well as the treatment with the inflammatory cocktail and the combination of both-increased gene and protein expression of SUCNR1 in epithelial cells, although the statistical significance was obtained in combination for gene expression and in the cocktail for protein expression (Figure 1a). As expected, the treatment with the inflammatory cocktail induced a significant increase in the expression of NLRP3, ASC and CASP1, while the treatment with succinate alone failed to significantly modify their expression (Figure 1c). ...
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... treatment with succinate-as well as the treatment with the inflammatory cocktail and the combination of both-increased gene and protein expression of SUCNR1 in epithelial cells, although the statistical significance was obtained in combination for gene expression and in the cocktail for protein expression (Figure 1a). As expected, the treatment with the inflammatory cocktail induced a significant increase in the expression of NLRP3, ASC and CASP1, while the treatment with succinate alone failed to significantly modify their expression (Figure 1c). The coadministration of succinate and the cocktail significantly potentiated the expression of NLRP3, IL1B and IL18 induced by the cocktail (Figure 1c Graphs show mRNA (n = 7) and protein (n = 3) expression of SUCNR1. ...
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... expected, the treatment with the inflammatory cocktail induced a significant increase in the expression of NLRP3, ASC and CASP1, while the treatment with succinate alone failed to significantly modify their expression (Figure 1c). The coadministration of succinate and the cocktail significantly potentiated the expression of NLRP3, IL1B and IL18 induced by the cocktail (Figure 1c Graphs show mRNA (n = 7) and protein (n = 3) expression of SUCNR1. Image of a representative Western Blot of one independent experiment. ...
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... analysis of the protein levels of pro-Caspase-1 revealed that the treatment with the cocktail induced a significant increase, whereas levels of this protein were significantly attenuated in siSUCNR1 cells treated with the cocktail and the combination (Figure 1d). The quantification of secreted IL-1β protein levels by ELISA showed a significant increase in HT29 cells treated with the inflammatory cocktail, which was potentiated with the combination of both cocktail and succinate. ...
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... quantification of secreted IL-1β protein levels by ELISA showed a significant increase in HT29 cells treated with the inflammatory cocktail, which was potentiated with the combination of both cocktail and succinate. A significant reduction in secreted levels of IL-1β was obtained in both experimental conditions via the silencing of SUCNR1 (Figure 1e). ...
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... as shown in Figure 1f, succinate levels were quantified and 130.4 ± 11.1 µM of this metabolite were detected in the supernatant of vehicle HT29 cells. The treatment with the cocktail failed to significantly modify levels of succinate which were also similar in siSUCNR1-transfected cells. ...