Fig. 4

NEDD4L mediates the biological effects of UBE2T. A UBE2T overexpression increased level of p-PI3K and p-AKT, which was blocked by simultaneous transfection of HA-NEDD4L. H1299 cells were transfected with vector, FlAG-UBE2T, or FlAG-UBE2T in combination with HA-NEDD4L, followed by IB with indicated Abs. B, C NEDD4L arrested growth-promoting phenotype induced by UBE2T overexpression. H1299 cells were transfected with vector, FlAG-UBE2T, or FlAG-UBE2T in combination with HA-NEDD4L, followed by cell proliferation assay (B) and colony formation assay (C). D NEDD4L depletion caused UBE2T accumulation to increase level of p-PI3K and p-AKT, which was abrogated by simultaneous UBE2T depletion. H358 cells were transfected with si-NEDD4L, si-NEDD4L in combination with si-UBE2T along with control RNAi, followed by IB with indicated Abs. E–F NEDD4L depletion stimulated cell progression, which was abrogated by simultaneous UBE2T depletion: H358 cells were transfected with siRNAs targeting NEDD4L alone or in combination with siRNA targeting UBE2T, along with scramble control, followed by cell proliferation assay (E) and colony formation assay (F). Shown are mean ± SEM, **P < 0.01, ***P < 0.001