The sumoylation pathway is up-regulated in MM. (A) Sumo-1 immunoblot of protein lysate from myeloma patient PCs. Total lysate was prepared from 105 cells and separated by electrophoresis on 5% to 20% gradient gels. (B) Sumo-1 immunoblot of total protein lysate from MMCLs. Total lysates were analyzed as in panel A. Samples loaded were normal B cell, normal PC, and then samples from the MMCLs, which were: 1, RPMI 8226; 2, U266; 3, ARP1; 4, ARD; 5, OPM1; 6, OPM2; 7, MM1.S; 8, MM1.R; 9, LR5; 10, KMS, 28BM; 11, KMS, 28PE; 12, NCI, H929; and 13, INA6. (C) Sumoylation pathway components on MM patient samples. Total lysate prepared from 105 cells was separated by electrophoresis on 10% to 20% gradient gels. Samples were probed with antisera to Ube2I, PIAS1, and ARF as indicated. (D) Sumoylation pathway components in total lysates were analyzed as in panel A. Samples loaded were normal B cell, normal PC, and then samples from the MMCLs, which were: 1, RPMI 8226; 2, U266; 3, ARP1; 4, ARD; 5, OPM1; 6, OPM2; 7, MM1.S; 8, MM1.R; 9, LR5; 10, KMS-28BM; 11, KMS-28PE; 12, NCI-H929; and 13, INA6. Samples were probed with antisera to Ube2I, PIAS1, Nse2, ARF, and histone 1, respectively. (E) Sumo-1 immunoblot of protein lysate from 3 normal B-cell samples and from 3 myeloma patient PC samples. Total lysate was prepared, separated by electrophoresis on 5% to 20% gradient gels, and probed with a Sumo-1 antibody.
