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Blood, Vol. 112, Issue 13, 5122-5129, December 15, 2008
Chronic lymphocytic leukemia antibodies with a common stereotypic rearrangement recognize nonmuscle myosin heavy chain IIA
Blood Chu et al.
112: 5122
Supplemental materials for: Chu et al
Files in this Data Supplement:
- Figure S1. CLL mAb with stereotypic IGHV1-69, IGHD3-16, and IGHJ3 rearrangements immunoprecipitate observable proteins (JPG, 85.1 KB)
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10% SDS-PAGE of the immunoprecipitate (IP) and supernatant (Sup) resulting from HEp-2 cell extracts treated with mAb having stereotypic IGHV1-69, IGHD3-16, and IGHJ3 rearrangements (CLL068 or CLL258) or not (CLL412). HEp-2 cell extract alone is also shown. Protein size (kDa) markers, mAb heavy chain (IgH) and light chain (IgL), as well as 225 and 45 kDa protein bands are indicated.
- Figure S2. Binding of CLL mAb with stereotypic IGHV1-69, IGHD3-16, and IGHJ3 rearrangements colocalizes with MYHIIA after disruption by ROCK inhibition (JPG, 172 KB)
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Untreated or Y27632 treated SNB19 cells were stained with CLL068 antibody (green) and (A) anti-MYHIIA antibody (red) or (B) phalloidin stained F-actin (red). Cells were visualized by confocal microscopy (original magnification ×600) separately and merged together. (A) CLL068 staining colocalizes with MYHIIA in untreated SNB19 cells, where MYHIIA appears in the nucleus as well as the cytoplasm (r = 0.77). Treatment with Y27632 disrupts MYHIIA cellular localization, with increased deposition in the membrane ruffle. CLL068 staining colocalizes completely with the Y27632 induced MYHIIA cellular redistribution (r = 0.82). (B) In contrast, CLL068 staining does not colocalize with F-actin (r = 0.48). F-actin stress fibers are also disrupted by Y27632 treatment, but CLL068 staining does not colocalize as well with redistributed F-actin (r = 0.63).
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