Fig. 7: USP28 is required to DYRK2 stabilization in response to DNA damage.
From: A novel feedback loop between DYRK2 and USP28 regulates cancer homeostasis and DNA damage signaling
A HEK-293T cells were transfected with Flag-DYRK2 and treated or not with Adryamicin/Doxorrubicin (ADR) (3 μg/mL) for 12 h. USP28 levels were determined by WB. B HeLa cells were transfected with indicated plasmids and treated with the ATR inhibitor VE-821 (2.5 μM) and/or Adryamycin (ADR) (3 μg/mL) during 14 and 12 h, respectively. Phosphorylation of CHK1 at S345 was used as control for the treatments. USP28 degradation by DYRK2 was determined by WB. C HEK-293T cells were transfected with the indicated plasmids in presence or absence of Flag-DYRK2 WT and treated or not with Adryamicin/ Doxorrubicin (ADR) (3 μg/mL) for 12 h. HA-USP28 degradation by Flag-DYRK2 was followed by WB analysis. D HEK-293T cells were transfected with Flag-USP28 WT or an inactive mutant (Flag-USP28 C171A) treated or not with ADR (3 μg/mL) for 12 h and DYRK2 endogenous stabilization was analyzed by WB. E HEK-293T cells were transfected with two different USP28 shRNAs and treated or not with ADR (3 μg/mL) for 12 h. Protein levels were determined by WB using specific antibodies. Note: All experiments were performed at least 3 times, and a representative blot is shown.
