Journal: Molecules

Article Title: Cell Reporter-Based Analysis of Anti-Inflammasome Activities of Flavonoids with Distinct Structures

doi: 10.3390/molecules31183176

Figure Lengend Snippet: Evaluation of the HEK-Blue IL-1R reporter platform for IL-1β detection. ( A ) Experimental protocol used to assess the dynamic range in response to IL-1β. ( B ) Statistical comparison among IL-1β standards ranging from 0.01 to 10,000 pg/mL as measured by the HEK-Blue IL-1R system. ( C ) Nonlinear four-parameter logistic (4PL) regression model generated from IL-1β standards. ( D ) Experimental workflow to obtain IL-1β-containing supernatants from BMMs in the presence or absence of LPS (10 ng/mL) and SEAP quantification using the HEK-Blue IL-1R assay. ( E ) Detection of SEAP activity over the course of a 5–15 min incubation with QUANTI-Blue reagent. N = 3, **** p < 0.0001, ns = not significant.

Article Snippet: SEAP levels, used as an indirect measure of IL-1β activity in the supernatant, were quantified using a colorimetric enzyme assay with QUANTI-Blue solution (InvivoGen, San Diego, CA, USA).

Techniques: Comparison, Generated, Activity Assay, Incubation

Journal: Molecules

Article Title: Cell Reporter-Based Analysis of Anti-Inflammasome Activities of Flavonoids with Distinct Structures

doi: 10.3390/molecules31183176

Figure Lengend Snippet: Quality controls for potential effects on cell viability and the Hek-Blue reporter platform. ( A , B ) LDH measurements in supernatants of BMM ( A ) and Hek-Blue cells ( B ) used in the experiments shown in . ( C ) Absorbance of flavonoid solutions (50 µM in water) measured at the indicated wavelengths (n = 3). ( D ) Hek-Blue cells were incubated with recombinant IL-1β (10 pg/mL, 24 h) in the presence of 25% media containing either no flavonoids or flavonoids at 10 or 50 µM. SEAP production was quantified using QUANTI-Blue (n = 4). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: SEAP levels, used as an indirect measure of IL-1β activity in the supernatant, were quantified using a colorimetric enzyme assay with QUANTI-Blue solution (InvivoGen, San Diego, CA, USA).

Techniques: Incubation, Recombinant