b-AP15isaspecificinhibitorofthedeubiquitinatingenzymesUch37andUsp14ofthe26Sproteasome.Itblocksthedeubiquitinatingactivityofthe26Sproteasome.Treatingcellswithb-AP15causestheaccumulationofpolyubiquitinatedproteins,inhibitionofproteasomaldegradation,upregulationofapoptoticmarkersandcelldeath.
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EqualamountofwholecelllysatespreparedfromDMSO(lane1)or2μMb-AP15(lane2)-treatedHeLacellswereseparatedbySDS-PAGEandimmunoblottedwithananti-Ubantibody.HEK293TCellsweretreatedwithDMSOorb-AP15for5hours.