Skip to main content
. Author manuscript; available in PMC: 2020 Sep 21.
Published in final edited form as: Cell Rep. 2020 Sep 8;32(10):108102. doi: 10.1016/j.celrep.2020.108102

Figure 1. SAG Competes with APC/C for UBE2C or UBE2S Binding to Affect APC/C Activity.

Figure 1.

(A–D) H1299 cells were transfected with siRNA targeting SAG or control siCont (A and B) or a plasmid expressing FLAG-tagged SAG or vector control (C and D) for 48 h with last 8-h treatment of 10 μM MG132 before harvesting. Cell lysates were prepared for immunoprecipitation (IP) with immunoglobulin G (IgG) control or antibodies (Abs) against UBE2C (A and C) or UBE2S (B and D), followed by immunoblot (IB) with indicated Abs. WCE, whole-cell extract.

(E and F) H1299 cells were transfected with siRNA targeting APC2 or control siCont for 48 h with last-8 h treatment of 10 μM MG132 before harvesting. Cell lysates were prepared for IP with IgG control or Abs against UBE2C (E) or UBE2S (F), followed by IB with indicated Abs.

(G–I) H1299 or H2170 cells were transfected with siRNA targeting SAG or control siCont for 48 h, followed by 24-h treatment of 12 uM pro-TAME (H) or 10 μM MG132 (I) in indicated samples before harvesting for IB analysis with indicated Abs.

(J–L) H1299 cells were transfected with various siRNA targeting indicated mRNAs, along with control siCont for 48 h. Cell lysates were prepared for IB analysis with indicated Abs.

Also see Figures S1 and S2.