A chemical approach for global protein knockdown from mice to non-human primates

a Mechanism of action of PROTAC. b Docking mode of RC32 binding to FKBP12 and recruiting CRBN. Moiety in red, linker; moiety in blue, rapamycin; moiety in green, pomalimide. c Chemical structure of RC32. d Immunoblots showing degradation of FKBP12 in Jurkat cells treated with RC32 at the indicated concentrations for 12？h. ？-Actin served as a loading control. e Immunoblots for FKBP12 protein. Jurkat cells were first treated for 3？h with bortezomib (Bortez), carfilzomib (Carf), rapamycin (RAP), or pomalimide (Poma) followed by treatment with RC32 (10？nM) for 2？h. ？-Actin served as a loading control. f DC50 with 12？h of RC32 treatment in Jurkat cells. g Recovery of FKBP12 level after washout of RC32. After treatment for 12？h with 1？μM RC32, the Jurkat cells were washed with fresh culture medium and further cultured in fresh medium for the indicated times. h Selectivity of RC32 against FKBPs in Jurkat cells. i Degradation efficiency of RC32 in different cell lines (100？nM) and primary cardiac myocytes (1？μM) treated for 12？h. Data in (f) and (i) are presented as mean？±？SEM of 3 independent experiment