Fig. 7
JOSD2 inhibitors abolish JOSD2 DUB activity. a Schematic illustration of JOSD2 inhibitors screening (top) and JOSD2 activity inhibition ratio of indicated compounds (n = 99) (bottom). Black dashed line indicates no inhibition on JOSD2 activity. 2 μM indicated compounds were incubated with GST-JOSD2 protein at 37 °C for 30 min and then subjected to in vitro ubiquitin-AMC assay. b Chemical structures of HY041004, HY04071 and HY032801. c 293FT cells transfected with LKB1-Flag, HA-Ub K6 and JOSD2-HA for 48 h were treated with indicated compounds for 4 h at concentration of 0.5 μM. IPs by Flag beads from cell lysates were subjected to IB. d 293T cells transfected with LKB1-Flag, STRAD-Myc, HA-Ub K6 and JOSD2-HA for 48 h were treated with indicated compounds for 4 h at concentration of 0.5 μM. IPs by Flag beads from cell lysates were subjected to IB. e PC-9 cells stably transfected with JOSD2-HA were treated with indicated compounds for 4 h at concentration of 0.5 μM and cell lysates were subjected to IB. f NCI-H358 cells were incubated with DMSO or 40 μM HY041004 and subjected to cellular thermal shift assay. Cell lysates were lysed by RIPA lysis buffer and then subjected to IB
