Figure 4

Gomesin peptides activate the Hippo pathway, inhibit MAPK cascades, and stimulate the p53/p21 cell cycle checkpoint axis. Representative western blots from three independent experiments showing: (a) Immunodetection of phospho-YAP^Ser127 in comparison to total YAP protein as a marker of the Hippo pathway; (b) Phospho-p42/44 (also called Phospho ERK) as a marker of the level of activation of the MAPK pathway; (c) PCNA, cyclin D1, p53, p21 and p27 as markers of the checkpoints that regulate progression of the proliferative cell cycle. Actin was used as a protein-loading marker; (d) Phospho-AKT^Ser473 in comparison to total AKT protein; (e) Phospho-Rictor^Thr1135 in comparison to total Rictor protein as a marker of the activation of the mTORC2 complex/cascade which is responsible for phosphorylation of AKT at Ser473; (f) Phospho-p70S6K^Thr389 in comparison to total p70S6K protein and Phospho-4E-BP1^Thr37/46 in comparison to total 4E-BP1 protein as markers of activation of the mTORC1 complex/cascade which is responsible for the phosphorylation of Rictor at Thr1135. MM96L cells were treated with 50 μg/ml AgGom (AgG) or HiGom (HiG) for 24 h. Right panel shows quantification of the western blots.