Anti aifm2

The cell extracts were prepared using RIPA buffer (KeyGen Biotech) containing protease inhibitors (KeyGen Biotech). Equal amounts of protein samples were subjected to 12% SDS-PAGE and transferred to PVDF membranes (Immobilon-P; Millipore, Billerica, USA). The membranes were then blotted with primary antibodies overnight at 4°C, followed by incubation with the HRP-conjugated goat anti-rabbit IgG secondary antibody (1:5000). The antibodies used were as follows: anti-AIFM2 (1:1000; Biorbyt, Cambridge, UK), anti-Bcl2 (1:2000; Abcam), anti-Bax (1:5000; Abcam); anti-Caspase3 (1:5000; Abcam), anti-Bak (1:1000; CST), anti-Cytc (1:1000; Abcam), anti-p53 (1:2000; Abcam), anti-p-mTOR (1:1000; CST), mTOR (1:1000; CST), anti-p-PI3K (1:1000; Affinity Biosciences), anti-PI3K (1:1000; Affinity Biosciences), anti-p-AKT (1:1000; CST), anti-AKT (1:1000; CST), anti-p-JAK2 (1:1000; Abcam), anti-JAK2 (1:1000; Abcam), anti-p-STAT3 (1:5000; Abcam), anti-STAT3 (1:5000; Abcam) and anti-GAPDH (1:5000; BBI, Shanghai, China). The protein bands were then visualized using enhanced chemiluminescence reagent (Bio-Rad, Hercules, USA). Band quantification was conducted using ImageJ (National Institutes of Health, Bethesda, USA).

Total protein was extracted from the mice pancreatic tissues. Primary antibodies used in this study include anti-ATF6 (Abcam; diluted 1: 500), anti-p53 (Abcam; diluted 1: 500), anti-AIFM2 (Biorbyt; diluted 1: 500), and anti-GAPDH (Abcam; diluted 1:1000). The protein abundance was evaluated by immunoblotting with at least three biological replicates. Please refer to the Supplemental Methods for more details.