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. Author manuscript; available in PMC: 2023 Mar 17.
Published in final edited form as: Cell Chem Biol. 2021 Nov 29;29(3):517–529.e5. doi: 10.1016/j.chembiol.2021.11.005

Fig. 5. p97 promotes the stability of cell-cycle oncoproteins.

Fig. 5

(a) qRT-PCR analysis of Myc, Securin, Emi and CDC20 mRNA levels. HCT116 cells were treated with 1 μM of MG132 or 2 μM of CB-5083 for 6h, n=4. (b-c) MG132 rescued CB-5083 mediated cyclin D1 downregulation at the protein level (b) but not at the mRNA level (c) while Baf A1 had no effect on cyclin D1. The concentration of MG132, CB-5083 and Baf A1 was 1 μM, 2 μM and 10 μM respectively. Cells were treated for 6 hours, n=4, **** indicates p<0.001. (d-e) The half-life of Securin in HCT116 cells. HCT116 cells were treated with 1 μM of MG132, 2 μM of CB-5083, 1 μM of MG132 plus 2 μM of CB-5083 or DMSO. 50 μM of CHX was added immediately after compounds treatment, n=3, * indicates p<0.05. (f-g) The degradation of Securin was detected in the total lysate of HCT116 and HT29 cells. Cells were pretreated with 2 μM of MG132 for 1h. Then harvest the cells as 0 minute samples, or replace the culture media with fresh media and add DMSO, 2 μM of CB-5083 or 1 μM of MG132 together with 50 μM of CHX for 60, 90, 120,180 minutes, n=3, * indicates p<0.05, ** p < 0.01. For fig. 5b–g, D, B, M, C and N represents DMSO, Baf A1, MG132, CB-5083 and NMS-873 respectively. Data are shown as mean ± SD. Statistical analysis was performed using one-way ANOVA. (See Fig. S5).