Fig. 4: F36V-VHL degrader outperforms CRBN and Halo targeting degraders.
a Western blot analysis of total cell lysates from immortalized MDFs (iMDFs) constitutively expressing H-E-N-F. Cells were stimulated with DMSO or stimulated with 500 nM of FC1, FC2, FV1, HV1, HV2 or NC4 degrader compounds for 5, 24 or 48 h. Asterisk (*) indicates possible cleavage product. b Western blot analysis of total cell lysates from immortalized MDFs (iMDFs) constitutively expressing H-E-N-F. Cells were stimulated with DMSO or stimulated with 500 nM of FC1, FC2, FV1, HV1, HV2 or NC4 degrader compounds for the first 5 h. Degraders were then removed from the culture medium at 5 h and cells were incubated for 24 or 48 h. Minus sign (−) indicates that sample was harvested at the time the treatment was removed (5 h). Asterisk (*) indicates possible cleavage product. c Schematic depicting the comparison of selected tTPD systems; F36V-VHL (FV1) and F36V-CRBN (FC1). d Western blot analysis of total cell lysates from MLKL−/− HT29 cells stably expressing the doxycycline-inducible C-terminal fusion protein MLKL-FKBPF36V. Cells were treated with 40 ng/mL doxycycline overnight to induce the constructs, then stimulated with the indicated concentrations of FV1 (F36V-VHL) or FC1 (F36V-CRBN) for 5 h. e Schematic depicting the comparison of selected tTPD systems; F36V-VHL (FV1) and Halo-VHL (HV2). f Western blot analysis of total cell lysates from MLKL−/− HT29 cells stably expressing the doxycycline-inducible C-terminal fusion protein MLKL-FKBPF36V or MLKL-Halo treated with 40 ng/mL doxycycline overnight to induce the construct, then stimulated with the indicated concentrations of FV1 (F36V-VHL) or HV2 (Halo-VHL). All experiments were repeated independently three times, and a representative Fig. is shown. Source data are provided as a Source Data file.
