Abin1

The T. gondii ME-49 strain was provided by the Laboratory of Parasitology, Jinan University. The Jurkat T-cells and Molt-4 T-cells were obtained from the American Type Culture Collection (ATCC). The rabbit monoclonal anti-human antibodies pNF-κBp65, A20, ABIN1, and Cleaved caspase-8 were from Cell Signalling Technology (Boston, USA), and the NF-κB p65, MALT1 antibody was from Abcam (Cambridge, UK). The mouse monoclonal anti-human GAPDH and secondary antibodies were purchased from Santa Cruz Biotechnology (Dallas, USA). shRNA for A20 and ABIN1 were designed and synthesised by Guangzhou OBIO Co, Ltd. (Guangzhou, China). CCK-8 kit was purchased from Dojindo (Kumamoto-ken, Japan). Nuclear and Cytoplasmic Protein Extraction Kit and Annexin V-FITC and propidium iodide (PI) kit were purchased from KeyGen Biotech (Nanjing, China). The TNIP1-3Flag-IRES2-EGFP recombinant plasmid was constructed by OBIO Co., Ltd. (Guangzhou, China). The MALT1 inhibitor MI-2 was purchased from MedChem Express (USA). The TUNEL Cell Apoptosis Detection Kit was purchased from Beyotime Biotechnology (Shanghai, China).

Proteins were extracted from the peri-infarct cortex using RIPA buffer (Beyotime, China). A cytoplasmic/nuclear protein extraction kit (Beyotime) was used to extract cytoplasmic and nuclear proteins. Western blot was performed as described previously [39 (link)]. Briefly, proteins were separated on 10% gels and transferred to PVDF membranes. After blocking with 5% skim milk, membranes were incubated with the primary antibodies at 4°C overnight. The following primary antibodies against specific proteins were used: ABIN1 (#4664, Cell Signaling Technology, USA, 1 : 1000), A20 (#5630, Cell Signaling Technology, USA, 1 : 1000), NF-κB p65 (#8242, Cell Signaling Technology, 1 : 1000), IκBα (18220-1-AP, Proteintech, USA, 1 : 000), phospho-IκBα (Ser32) (#2859, Cell Signaling Technology, 1 : 1000), GAPDH (10494-1-AP, Proteintech, USA, 1 : 1000), and histone H3 (#3638, Cell Signaling Technology, 1 : 1000). Membranes were washed with TBST and immersed in the secondary antibodies at 37°C for 1 h. After washes with TBST, immunoreactive bands were detected using WesternBright ECL (Advansta, USA). Membranes were scanned and analyzed using a Fusion FX5 analysis system (Vilber Lourmat Fusion FX 7 Spectra, France).