Raw Western Blot Data for Protein X Expression in Condition Y

The primary antibodies used were as follows: FBXW11 (A7784, ABclonal), HIC1 (24949-1-AP, Proteintech), IRF1 (A7692, ABclonal), PARP (9542, CST), cleaved PARP (#5625, CST), caspase-3 (9662, CST), cleaved caspase-3 (9661, CST), IL-1β (12703, CST), cleaved IL-1β (83186, CST), Actin (AC004, ABclonal), and Anti-Ubiquitin (ab7780, Abcam). The membranes were washed three times with TBST, each for 10 minutes, followed by incubation with horseradish peroxidase (HRP)-conjugated secondary antibody (goat anti-rabbit IgG H&L, HRP, ab97051, 1:2000, Abcam, Cambridge, UK) for 1 hour. After TBST washes, the membranes were placed on a clean glass plate. Pierce™ ECL detection reagent (product number 32209, Thermo) was prepared by mixing equal volumes of solutions A and B in the dark, and the mixture was applied to the membranes. Imaging was performed using the Bio-Rad imaging system (ChemiDoc™ XRS+, BIO-RAD).

For the WB experiment, total proteins were extracted from cells using RIPA lysis buffer containing PMSF (P0013B, Beyotime, Shanghai). Protein concentrations were determined with a BCA Protein Assay Kit (T23225, Thermo Fisher Scientific, Rockford, IL, USA). 50 μg of protein was dissolved in 2× SDS loading buffer, boiled at 100°C for 5 minutes, and then subjected to SDS-PAGE for electrophoresis. The proteins were wet-transferred onto a PVDF membrane, followed by blocking with 5% non-fat milk at room temperature for 1 hour. The membrane was then incubated overnight at 4°C with primary antibodies. The primary antibodies used included: FBXW11 (ABCLONAL, A7784), HIC1 (Proteintech, 24949-1-AP), IRF1 (ABCLONAL, A7692), PARP (CST, 9542), cleaved PARP (CST, 5625), caspase-3 (CST, 9662), cleaved caspase-3 (CST, 9661), IL-1β (CST, 12703), cleaved IL-1β (CST, 83186), Actin (ABCLONAL, AC004), and Anti-Ubiquitin (Abcam, ab7780). After primary antibody incubation, the membrane was washed thrice with TBST, each lasting 10 minutes. The membrane was then incubated with an HRP-conjugated secondary antibody (goat anti-rabbit IgG H&L, HRP, Abcam, ab97051, 1:2000) for 1 hour. For detection, the membrane was treated with the Pierce™ ECL detection reagent (product number 32209, Thermo) by mixing equal volumes of solutions A and B and applying the mixture onto the membrane. Imaging was performed using a Bio-Rad imaging system (ChemiDoc™ XRS+, BIO-RAD).