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. 2020 Jan 2;11(1):5. doi: 10.1038/s41419-019-2199-4

Fig. 5. The acceleration of MCL1 degradation by ALG-2 relies on its interaction with Rpn3.

Fig. 5

a The expression level of Rpn3 in Rpn3-KO Jurkat cell line. The protein levels of Rpn3 and GAPDH are determined by western blotting. b The proliferation rate of Rpn3-KO Jurkat cells. The assay was started with 500,000 cells and measured by Countstar cell-counter system. Data shown were representative of three independent experiments. c The proteasome activity in Rpn3-KO cell line was impaired, but was rescued with re-expression of Rpn3. d, e ALG-2 loses the ability to accelerate MCL1 degradation in Rpn3-KO Jurkat cell line. Rpn3-KO Jurkat cells were infected with ALG-2 retrovirus (d) or sh-ALG-2 retrovirus (e) for 40 h, then treated with CHX and MG132 for another 8 h. Cells were collected and lysed in RIPA buffer for detection. f The effect of ALG-2 on the stability of MCL1 when Rpn3 was rescued in Rpn3-KO cell. Data shown were representative of three independent experiments. Error bars indicate SD. ns, no significance; ***P < 0.001.