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Fig. 5 | Journal of Inflammation

Fig. 5

From: Acidification changes affect the inflammasome in human nucleus pulposus cells

Fig. 5

Exogenous IL-1β contributes to the inflammatory response in HNPC. Cells were grown at 3 different pH levels (7.4, 6.8, 6.5) for 24 h and the media was used to run a lactose dehydrogenase (LDH) release assay. Data presented as mean+/−SEM. **p < 0.05 compared to 6.8 and *p < 0.05 compared to 7.4. N = 6 wells per group (a). Cells were then grown at a pH of 6.5 and treated with human recombinant protein to IL-1β at different concentrations (1, 5, 10 ng/ml) (b). Representative immunoblot analysis of ASC protein expression in HNPC treated with different concentrations of IL-1β (1, 5, 10 ng/ml) for 24 h at a pH of 6.5 (c). β-actin was used as an internal standard for protein loading control. Data presented as mean+/−SEM. *p < 0.05 compared to control. N = 6 wells per group. Silencing of ASC/pycard by siRNA in HNPC resulted in increased caspase-1 and NLRP1 protein expression as well as increased XIAP cleavage when cells were treated with 10 ng/ml of human recombinant IL-1β protein at a pH of 6.5. (+) indicates ASC/pycard was not silenced and (−) indicates ASC was silenced (d). Representative model of the effects of ASC silencing and exposure to IL-1β on inflammasome activation in HNPC (e)

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