Fig. 4

A High-throughput phosphorylation profiling with 1318 site-specific antibodies from over 30 signaling pathways was performed in the lysates of MINK1KO and MINK1WT clones of H357 5FUR cells as described in methods. The top 20 upregulated phosphoproteins (MINK1 KO/MINK1WT) are represented in left panel, whereas top 20 downregulated phosphorylated proteins are represented in right panel. The upregulated targets considered in the study is marked in red box, whereas downregulated targets in green box. B Lysates were collected from indicated cells and immunoblotting (n = 3) was performed with indicated antibodies. C pLenti CMV/TO Puro DEST MINK1 (ectopic overexpression of MINK1) was transiently transfected in 5FUR lines stably expressing MINK1 ShRNA (targeting 3′UTR) and immunoblotting (n = 3) was performed with indicated antibodies. D Lysates were collected from indicated cells and immunoblotting (n = 3) was performed with indicated antibodies. E MINK1 was ectopically overexpressed in 5FUR lines stably expressing MINK1 ShRNA (targeting 3′UTR) and immunoblotting (n = 3) was performed with indicated antibodies. F pLNCX myr HA Akt1 (ectopic overexpression of myr AKT) was transiently transfected in indicated MINK1KO cells and immunoblotting (n = 3) was performed with indicated antibodies. G MINK1 was ectopically overexpressed in 5FUR lines stably expressing MINK1ShRNA (UTRKD) as described in panel C and treated with AKT inhibitor (Akti-1/2) for 24 h and immunoblotting (n = 3) was performed with indicated antibodies. H Lysates were collected from indicated cells and immunoblotting (n = 3) was performed with indicated antibodies.