Phosphorylated nf κb p65 s536

The western blotting assay was performed as previously described²² . The following primary antibodies were used: β-actin (Proteintech, 60008-1-Ig), Vimentin (Cell Signaling Technology, 5741), N-cadherin (Cell Signaling Technology, 13116), β-Catenin (Cell Signaling Technology, 8480), CD44 (Proteintech, 15675-1-AP), MMP14 (abcam, ab51047), Phosphorylated NF-κB p65 (S536) (Cell Signaling Technology, 3033), NF-κB p65 (Cell Signaling Technology, 8242), and HIF-1α (Cell Signaling Technology, 36169).

Whole-cell protein was extracted from glioma cells and MDSCs in RIPA buffer (Thermo Fisher Scientific, USA) and centrifuged at 12,000 rpm for 20 min. A BCA kit (Thermo, Waltham, MA, 23228) was used to measure the protein concentration. After immunoblotting, the proteins were transferred to a nitrocellulose membrane and incubated with specific antibodies. The following primary antibodies were used: β-actin (Proteintech, 60008-1-Ig), CyclinD1 (Cell Signaling Technology, 2978), P27 (Cell Signaling Technology, 3686), CDK6 (Cell Signaling Technology, 3136), p-AKT (Cell Signaling Technology, 4060), AKT (Cell Signaling Technology, 4691), SETD7 (Cell Signaling Technology, 2813), hnRNPA2/B1 (Cell Signaling Technology, 9304), Phosphorylated NF-κB p65 (S536) (Cell Signaling Technology, 3033), and NF-κB p65 (Cell Signaling Technology, 8242).