Fig. 7

Uev1A regulates CT45A expression through the AKT signaling pathway. a The cellular AKT protein and its phosphorylation (p-AKT-Thr308, p-AKT-Ser473) levels in pcDNA4.0/TO/HA (+) vector (CK), UEV1A, UEV1A-F38E transiently transfected MDA-MB-231 (left panel) and MCF7 (right panel) cells were monitored by western blot using anti-AKT, anti-p-AKT-Thr308 and anti-p-AKT-Ser473 antibodies. b The UEV1A transiently transfected MDA-MB-231 (left panel) and MCF7 (right panel) cells were treated with 10 μM PI3K/AKT pathway inhibitor LY294002. After 24 h, the AKT and p-AKT-Ser473 levels were examined by western blot using anti-AKT, anti-p-Ser473 antibodies in cells transfected with vector, UEV1A with or without LY294002 treatment as indicated. Ectopic UEV1A expression was detected by an anti-HA antibody. c, d Relative CT45A expression levels in MDA-MB-231 (c) and MCF7 (d) cells transfected with vector, UEV1A with or without LY294002 treatment as indicated, followed by qRT-PCR. e, f MDA-MB-231 (e) and MCF7 (f) cells were treated with IGF-1 over time as indicated and the cellular AKT and p-AKT-Ser473 proteins were monitored by western blot using anti-AKT and anti-p-AKT-Ser473 antibodies. g, h Transcript levels of CT45A in MDA-MB-231 (g) and MCF7 (h) cells treated with IGF-1 over time were monitored by qRT-PCR. CK, control treatment; LY, LY294002. All experiments were performed in at least triplicate and the results are the average with standard deviation. **, P < 0.01. a, b, e, f The gel images in are cropped from available original blots and numbers underneath the WB images indicate relative band intensity after normalization with the loading control