Journal: SLAS discovery : advancing life sciences R & D
Article Title: Discovery and validation of small molecule stabilizers of mutant triose phosphate isomerase (TPI) as potential lead candidates for TPI deficiency
doi: 10.1016/j.slasd.2025.100278
Figure Lengend Snippet: Appearance and behavior of the HeLa TPI E105D-GF-HR cell line. Upper panel. Representative images of cells expressing mutant TPI-GFP (green) and a HyPerRed ROS biosensor (orange). Nuclei were counterstained with Hoechst 33342 (blue). Images were acquired on an OPERA Phenix Plus using a 63x water immersion objective in the DAPI, GFP, and Cy3 channel. Lower panel. Quantification of cellular response to test agents reveals desirable (elevated GFP in the absence of nuclear shrinkage and elevated ROS; luminespib) and undesirable activity profiles (elevated GFP correlating with nuclear shrinkage and elevated ROS; CID 3724239). Y-axes show percent increases compared with vehicle-treated cells. Data points are the average of at least three independent biological repeats ± SD. Curves were fitted to a four parameter logistic regression. Green, GFP-TPI levels, red, ROS, blue, nucleus size. Images were subjected to identical linear bitmap stretch for visibility.
Article Snippet: A construct with the EF1 promoter driving HyperRed protein expression was generated using a HyperRed clone (Addgene # 48,249 [ ], and standard cloning techniques.
Techniques: Expressing, Mutagenesis, Activity Assay