Fig. 3: Bioactivity evaluation of aMD4dY-PA22.
From: Single-chain tandem macrocyclic peptides as a scaffold for growth factor and cytokine mimetics
a Phosphorylation of Met induced by aMD4dY-PA22. Met phosphorylation (Y1234/Y1235) level of HuCCT1 cells stimulated using aMD4dY-PA22 (red squares), rhHGF (blue circles), and monomeric aMD4dY (green triangles) was evaluated via phosphor-Met ELISA. The mean and individual values are shown from the results of triplicate experiments. b Signaling potencies induced by aMD4dY-PA22. Erk signaling activation by aMD4dY-PA22 (red squares), rhHGF (blue circles), and monomeric aMD4dY (green triangles) was evaluated by Erk-SRE reporter assay. The mean and individual values are shown from the results of triplicate experiments. c Selectivity of aMD4dY-PA22 against various RTKs. Cell lysates of HEK293 cells stimulated by 1.3 nM rhHGF or 32 nM aMD4dY for 10 min were analyzed by a phospho-RTK array. Positions of phospho-Met are indicated by red boxes. d Wound healing promoted by aMD4dY-PA22. Wound closure events of HuCCT-1 cell monolayer at 1 and 9 h after scratching with or without 0.41 nM rhHGF or 16 nM aMD4dY-PA22 were captured. Scratches are depicted by red broken lines. Scale bars = 1 mm. Average wound closure rates from 1 to 7 h of each experimental group were calculated. The mean ± SD with individual values are shown from the results of quadruplicate experiments. ****p < 0.0001. e Branching morphogenesis induced by aMD4dY-PA22. HuCCT-1 cells were encapsulated in collagen gels and cultured in the presence of 0.44 nM rhHGF or 6.3 nM aMD4dY-PA22 for 7 days. Arrows indicate tubule structures. Scale bars = 200 μm.
