Supplementary Figure S6 from TR-107, an Agonist of Caseinolytic Peptidase Proteolytic Subunit, Disrupts Mitochondrial Metabolism and Inhibits the Growth of Human Colorectal Cancer Cells
Supplementary Figure S6. Washout of TR-107 treatment suggests an irreversible inhibition of CRC cell viability. (A) HCT116 cells were treated in a 96-well plate format with vehicle control (0.1% DMSO) or indicated concentrations of TR-107 for 2, 4, 6, 8, 12, 24, and 72 hours. At each time point, wells were washed out twice with fresh media and replaced with fresh media lacking treatment up until 72 hours. Cell viability was assessed for all treatment durations after 72 hours. Data are presented as mean ± SD values (N=6). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (B) Western blot analysis of HCT116 cells after 24 hours of treatment with washout and 72 hours with no washout. HCT116 cells were treated with vehicle control (0.1% DMSO) or TR-107 10 nmol/L, 50 nmol/L, 1 μmol/L. Immunoblot was performed for ClpX, mtTFA, and β-actin as loading control.