Fig. 4: PI3K signaling enhances AKT1-R391 dimethylation.
From: PRMT5-mediated arginine methylation activates AKT kinase to govern tumorigenesis
a, b IB analysis of WCLs and IP products derived from MCF7 cells. The cells were serum-starved for 16 h and then treated with 100 nM insulin for indicated time periods before collection. c IB analysis of WCLs and IP products derived from MCF7 cells. The cells were serum-starved for 24 h and treated with PI3K inhibitors for 12 h followed by insulin for 30 min before collection. LY294002 (20 μM), Wortmannin (10 μM), and Buparlisib (10 μM). d, e IB analysis of WCLs and IP products derived from HEK293T cells transfected with indicated constructs. f In vitro binding assays were performed with recombinant GST-AKT1 protein purified from mammalian cells and Myc beads bound with PRMT5/MEP50. The binding was performed at 4 °C for 4 h incubated with or without PIP3 (20 μM) and subjected to IB analysis. Similar results were obtained in n ≥ 2 independent experiments in a–f. Uncropped immunoblots are provided in Source Data file.
