Hsp90 Isoform-Specific Inhibitor Synthesis

The inhibitors for specific Hsp90 isoforms [KUNA115 and NDNA1065 (Hsp90α), KUNB106 and its analog, NDNB1151 (Hsp90β), KUNG65 (Grp94), T1, T2, and T3 (TRAP1)] were synthesized and purified by the laboratory of Dr. Brian Blagg at the University of Notre Dame [(Crowley et al., 2017 (link); Mishra et al., 2021a (link); Mishra et al., 2021b (link); Merfeld et al., 2023 (link)), Figure 1]. Lipopolysaccharide (LPS, from E. coli O111:B4), 17-AAG and penicillin:streptomycin (tissue culture) were purchased from VWR (Radnor, PA, United States). Dulbecco’s Modified Eagle Medium (DMEM), trypsin-EDTA, fetal bovine serum (FBS), and G418 disulfate solution were obtained from Corning (Corning, NY, United States). Griess reagents for nitric oxide assay (sodium nitrite, sulfanilamide, N-1-naphthylethylenediamine) were purchased from Sigma-Aldrich (St. Louis, MO, United States). Antibodies used for Western blots include goat anti-rabbit IgG-horseradish peroxidase, and goat anti-mouse IgG-horseradish peroxidase (Invitrogen, Waltham, MA, United States); p-ERK, t-ERK, p-p38, t-p38, p-JNK, and t-JNK (Cell Signaling, Beverly, MA, United States).