Rat il 1β

Rat IL-1β (Interleukin 1 Beta) ELISA Kit (E-EL-R0012c), IL-6 (Interleukin 6) ELISA Kit (E-EL-M0044c), IL-17 (Interleukin 17) ELISA Kit (E-EL-R0566c), TNF-α (Tumor Necrosis Factor Alpha) ELISA Kit (E-EL-R2856c), IFN-γ (interferon-γ)ELISA Kits (E-EL-R0009c) were purchased from Elabscience Biotechnology.

Antibodies were as follows: GSDMD antibody (Lot: #E9S1X) was purchased from Cell Signaling Technology (Massachusetts, USA). Caspase-1 (Lot: #ab179515) and S100 antibodies (Lot: #ab52642) were purchased from Abcam (Cambridge, England, UK). NLRP3 (Lot: # T55651) and Rab32 (Lot: # TD9825) was purchased from Abmart (Shanghai, China).
Reagents were as follows: Rat IL-1β (Lot: #E-EL-R0012c) and IL-18 (Lot: #E-EL-R0567c) ELISA Kits were obtained from Elabscience (Shanghai, China). Lipopolysaccharides (LPS, Lot: #L4391-1MG) and adenosine triphosphate disodium (ATP, Lot: #A2383-5G) was purchased from Sigma-Aldrich. We obtained Mitoquinone mesylate (MitoQ, Lot: # HY-100116A) from MedChemExpress (Monmouth Junction, NJ, USA).

In order to analyze the evolution of the markers of inflammation, the plasma was separated from the rest of the blood of the samples, by means of the centrifugation technique at 1500 rpm during 10’ at 4 °C. Once the plasma samples were separated, three ELISA tests were performed to determine the concentrations of the different interleukins studied:
IL-6, IL-1β Y TGF-β1, according to their respective manufacturer protocols: Rat IL-6 ELISA KIT of Diaclone^® (Besançon, France), Rat Transforming Growth Factor 1β (TGF- 1β) ELISA KIT of Cusabio^® (Houston (TX), USA), and Rat IL-1β (Interleukin 1 β) ELISA KIT of Elabscience^® (Houston (TX), USA). The CLARIOstarplus system and the MARS^® Software, both of BMG LABTECH© (Ortenberg, Germany), were used to read the plates.