Anti prdx4

For in vitro experiments, total protein isolation was performed using the RIPA lysis buffer according to the standard techniques. The mitochondria were removed, and the cytoplasmic protein was extracted using the Cell Mitochondria Isolation Kit (Beyotime) according to the manufacturer’s instruction. For in vivo experiments, the xenograft tissue was flash-frozen in liquid nitrogen and carefully ground, and then the tissue was lysed in RIPA lysis buffer.
Western blot analysis was performed as previously described.^{15 (link)} The following primary antibodies were used: anti-IDH2 (1:1000; Proteintech, Chicago, IL, USA), anti-IDH1 antibody (1:1000; Proteintech), anti-GAPDH (1:4000; Proteintech), anti-PRDX1 (1:10000; Abcam, Cambridge, UK), anti-PRDX2 (1:1000; Abcam), anti-PRDX3 (1:10000; Abcam, anti-PRDX4 (1:10,000; Abcam), anti-PRDX5 (1:1000; Abcam), anti-TXN (1:500; Abcam), anti-TXN2 (1:10,000; Abcam), anti-SOD2 (1:1000; Abcam), anti-DRP1 (1:1000; Abcam), anti-OPA1 (1:1000; Abcam), anti-Phospho-AKT (pAKT) (T308) [1:1000; Cell Signaling Technology (CST), Danvers, MA, USA], anti-Bax (1:1000; CST), anti-Bcl-2 (1:1000; CST), anti-cytochrome c (1:1000; CST), anti-cleaved caspase-3 (1:1000; CST), and β-actin (1:2000; Proteintech).