Skip to main content
. 2016 Jun 15;6:28016. doi: 10.1038/srep28016

Figure 4. Chac1 (K0) devoid of lysines is still ubiquitinated in HEK293 cells.

Figure 4

(a) HEK293 cells were transiently co-transfected with 5′-Chac1 (WT or K0)-Myc and HA-Ub (WT). After 24 h, the cells were treated with vehicle (con) or 20 μM MG for 12 h, and the resultant cell lysates were analyzed by western blot using c-Myc or Actin antibodies. (b) HEK293 cells were transiently co-transfected with Chac1 (WT or K0)-MycHis and HA-Ub (WT). After 24 h, the cells were treated with 20 μM MG for 12 h. Myc-tagged proteins were immunoprecipitated from the resultant cell lysates using a c-Myc antibody and Protein G Sepharose and were analyzed by western blot using HA or c-Myc antibodies. The total cell lysate (Input) was analyzed by western blot using c-Myc antibody. Figure 4a,b data were obtained from three and two independent cultures, respectively. Ub-Chac1 indicates the ubiquitinated Chac1.