Skip to main content
. 2014 Jun;34(12):2264–2282. doi: 10.1128/MCB.01542-13

FIG 4.

FIG 4

COPS5 stabilizes PEA15 protein by deubiquitination. (A) Immunoblot of PEA15 and p27 in HCT116 or HeLa cells treated with indicated concentrations of MLN-4924 under unirradiated or gamma-irradiated conditions. Actin is used as the loading control. (B) RT-qPCR analysis to measure mRNA levels (n = 3) of COPS1, COPS3, COPS6, and USP15 in HCT116 cells stably expressing an NS shRNA or two different shRNAs (1 and 2) against each of the indicated genes. Error bars indicate standard errors of the means. (C) Immunoblot analysis of PEA15 in HCT116 cells carrying the indicated shRNAs under unirradiated (left) or gamma-irradiated (γ-rad) conditions (right). Actin is used as the loading control. (D) Immunoblot analysis of polyubiquitinated PEA15 probed with the antibody against HA incubated without or with immunoprecipitated COPS5 for the indicated time. (E) Immunoblot analysis of polyubiquitinated PEA15 probed with the antibody against HA incubated without or with immunoprecipitated COPS5 or COPS5-D151N for 30 min. (F) Immunoblot analysis of polyubiquitinated PEA15 probed with the antibody against HA that was incubated with purified GST or GST–COPS5-WT or GST–COPS5-D151N for 2 h in the absence or presence of NEM or ubiquitin (Ub) aldehyde.