Fig 2: The CRBN65 antibody detects CRBN protein across a wide range of MM cell lines with high specificity. A panel of MM cell extracts (20 µg/lane) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and probed with CRBN65 antibody (1:1000) or ß-tubulin, without blocking peptide (upper panel) or with blocking peptide (lower panel). CRBN, cereblon; kDa, kiloDalton; MM, multiple myeloma; rCRBN, recombinant cereblon.

Fig 3: A schematic of CRBN splice variants crbncc-001 to crbncc-022 sequenced from MM cell lines with regions recognized by CRBN65 and TaqMan denoted. CRBN, cereblon; DDB1, DNA damage binding protein 1; IMiD, IMiD immunomodulatory drug; THAL, thalidomide.

Fig 4: CRBN mRNA and CRBN protein levels do not correlate in MM cell lines with intrinsic sensitivity or resistance to the IMiD agent lenalidomide. CRBN mRNA and CRBN protein levels were measured in 12 human MM cell lines by TaqMan polymerase chain reaction and Western blot, respectively. mRNA and protein were normalized to HRPT1 and ß-tubulin, respectively. Graph represents the average of 3–4 experiments. CRBN, cereblon; LEN, lenalidomide; MM, multiple myeloma; mRNA, messenger RNA.

Fig 5: The CRBN65 antibody detects CRBN protein across a range of normal human cells and tumour cells. A panel of cell extracts (20 µg/lane) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and probed with CRBN65 antibody (1:1000) or actin antibody. Cell types of cell lines tested are as follows: KMS11 and KMS34, human multiple myeloma; Rec1, mantle cell lymphoma; OCI-LY10, activated B-cell–like diffuse large B-cell lymphoma; Jurkat, T-cell leukaemia; HS5, bone marrow stroma fibroblast; NHNP, normal human progenitor cells; G166, glioblastoma; Br231 and Br468, breast cancer cell lines; PC3, prostate carcinoma cell line; LNCAP, prostate adenocarcinoma cell line; CRBN, cereblon; kDa, kiloDalton.