Figure 1.

General workflow for studying the effects of novel chalcone derivatives CH-1, CH-2, and CH-3 on TNBC cell lines and non-tumor breast cell lines. Potency assays (viability and clonogenic assays) were performed for all cell lines with each derivative. Each cell line was then treated with DMSO (control) or 5 μM derivative for 12 hours before harvesting and labeling with TMT labels. Each TMT six-plex experiment contained one cell line treated with each derivative and DMSO, and contained two biological replicates of the DMSO-treated and CH-2-treated cells. The samples were analyzed by LC/MS-MS and processed using MaxQuant, Perseus, and GSEA in order to determine the main proteins, pathways, and processes affected by these derivatives. Validation in the form of cell cycle, apoptosis, and Western blot assays were then performed.