Extended Data Fig. 3: ILF2 interacts with ILF3 and acts as a negative regulator of amino acid-stimulated mTORC1 activity.
From: Genome-wide CRISPR screens identify ILF3 as a mediator of mTORC1-dependent amino acid sensing
a, HEK293T cells transfected with the indicated cDNAs were starved of amino acids for 50 min, or starved and re-stimulated with amino acids for 10 min. Cell lysates were immunoprecipitated with FLAG beads. The whole-cell lysates (WCLs) and IPs were immunoblotted to detect the indicated proteins and phospho-proteins. b, HEK293T cells stably expressing Flag-tagged Metap2 or ILF3i1 were treated as in a, and then subjected to immunoprecipitation with FLAG beads. The WCLs and IPs were immunoblotted to detect the indicated proteins and phospho-proteins. c, Cells expressing Ctrl or ILF3 shRNAs were infected with lentivirus to co-express sgRNAs targeting GFP or ILF3. Cells were then treated as in a, and immunoblotted to detect the indicated proteins and phospho-proteins. d, HEK293T cells were transfected with S6K1 cDNA combined with the indicated cDNAs, treated as in a, and immunoblotted to detect the indicated proteins and phospho-proteins. Unprocessed blots are available in source data.
