Fig 1: Schematic diagram showing the anti-tumor mechanisms of CVM-1118 via targeting TRAP1. Molecular mechanisms in tumor cells to show how TRAP1 mediates anti-apoptosis and promotes VM formation (A) and the effects of CVM-1118 treatment (B).
Fig 2: CVM-1125 binds strongly to TRAP1 (A) To identify CVM-1125 binding target, the NPOT assay was performed on CVM-1125 treated MCF7 and COLO205 cells, as well as the colorectal and melanoma tumor homogenates. The experiments were performed three times independently. Arrows point at the isolated drug specific macromolecular assemblies. (B) The binding between TRAP1 and CVM-1125 was confirmed using SPR. There is a dose-dependent interaction between CVM-1125 and the immobilized TRAP1 with K D of 6.7 × 10−8 M. However, no specific interaction of Hsp90 to CVM-1125 was observed. (C) In silico interaction analysis of CVM-1125 with TRAP1 at the N-terminal domain (NTD).
Fig 3: Changes of TRAP1, HIF-1α and succinate levels after CVM-1125 treatment (A) TRAP1 protein levels, with or without CVM-1125 treatment, in COLO205 and SKOV3 cell lines. The protein level of TRAP1 was examined by Western blot and the relative expression at different treatment concentration was calculated by normalization to β-actin and normal control (NC) set as 1.0. Data represent median ± interquartile range for not normally distributed data in COLO205 cells and mean ± SD for normally distributed data in SKOV3 cells. (B) TRAP1 protein levels after treatment with CVM-1125 and chloroquine (CQ) lysosome inhibitor or proteasome inhibitor (MG132) in COLO205 cell line. Relative TRAP1 protein level was calculated by normalization to β-actin and DMSO vehicle control set as 1.0 and is shown in right. Significant difference (* = p < 0.05) is shown in the CQ treatment group that rescues the inhibition of TRAP1 expression by CVM-1125 treatment alone (n = 3). (C) Succinate levels without (Vehicle control) or with CVM-1125 treatment in SK-MEL28, MDA-MB-231, COLO205, and SKOV3 cell lines. (D) HIF-1α protein levels without or with CVM-1125 treatment in SK-MEL28, MDA-MB-231, COLO205, and SKOV3 cell lines. The relative amount of HIF-1α protein level was calculated against the untreated control cells normalized to a value of 1.0 after correcting for loading of β-actin protein measured in the control lane.
Supplier Page from Abcam for Recombinant human TRAP1 protein