Il 17a

The murine liver and ileum were fixed in 4% paraformaldehyde overnight at 4°C, dehydrated, soaked in xylene, embedded in paraffin in sequence, and then sliced into 4-μm sections. Paraffin sections were dewaxed with xylene and then dehydrated with different concentrations of ethanol. Sections were subjected to H&E staining and immunohistochemical as well as immunofluorescence staining. The following primary antibodies were used for immunohistochemical staining: IL-1β (Bioss, China), IL-6 (Bioss), IFN-γ (Bioss), IL-17A (Bioss), Caspase-1 (ABclonal, China) and NLRP3 (ABclonal). For immunofluorescence on tissues, ZO-1 (Abcam, UK), occludin (Abcam), F4/80 (Abcam), and Alexa Fluor 555 (BBI, China) were used. Furthermore, images were obtained under a microscope (AxioObser Z1, Germany) at a magnification of ×200, and positive results were quantified using ImageJ software (Free Software Foundation Inc., Boston, MA).

The amounts of IL-5, IL-13 (Th2 cytokines) and IL-17 A (Th17 cytokine) were assessed by specific enzyme-linked immunosorbent assay (ELISA) kits (Bioss Inc., China) as directed by the manufacturer. The relative mRNA levels of cytokines in HBE cells were assessed by qRT-PCR. The protein expression levels of IL-17 A (Invitrogen, USA), HDAC2 (Invitrogen, USA) and RORt (Invitrogen, USA) were detected by Western blotting and immunofluorescence analysis. Flow cytometry antibodies detecting HDAC2, RORt and IL-17 A were provided by Abcam (US). Flow cytometry was carried out on a BD Calibur instrument (BD, USA).