Dysregulated TCL1 requires the germinal center and genome instability for mature B-cell transformation -- Shen et al. 108 (6): 1991 Data Supplement - Supplemental materials for: Shen et al -- Blood

Dysregulated TCL1 requires the germinal center and genome instability for mature B-cell transformation
Blood Shen et al. 108: 1991

Supplemental materials for: Shen et al

Files in this Data Supplement:

Figure S1. A similar proliferation advantage for TCL1-tg GC and non-GC B cells (79.1 KB) -
Isolated TCL1-tg and WT spleen B cells were incubated with CFSE, stimulated with anti-IgM plus IL-4 and stained for B220 and PNA-receptor surface expression. Live-gated B220+PNA— non-GC B cells or B220+PNAhi GC B cells and those that acquire a GC phenotype from in vitro stimulation were evaluated for CFSE content. The percentage of viable cells with 3 or more cell divisions is listed for each population. The data are representative of 3 separate experiments.
Figure S2. Histology and flow phenotype of TCL1-tg tumors (PDF, 75.7 KB) -
(A) Hematoxylin and eosin stain (left panel) of spleen with histiocytic infiltrate and immunostain for PAX5 expression (right panel) in clonal, IgM+B220+CD5+PAX5+ B cell lymphoma. (B) Flow cytometry of spleen cells from a moribund 11-month-old TCL1-tg mouse (T5, Table S2) shows a monomorphic population of IgM+B220+CD5+ B cells that is identical to GC tumors developing in TCL1-tg mice described previously (12).
Figure S3. Clonality of lymphomas in TCL1-tg mice (PDF, 52.8 KB) -
Intense 4.7kb and 6.5kb bands represent StuI and EcoRI digests of germline IgH, respectively. Genomic DNA was digested with either StuI or EcoRI from spleen B cell lymphomas (lanes 2 and 4) or from histiocytic sarcoma cells (lanes 1 and 3) of TCL1-tg mice.
Figure S4. Lack of TCL1 expression in splenic macrophages of TCL1-tg mice (PDF, 21.5 KB) -
Cells isolated by magnetic bead sorting were immunoblotted for TCL1 expression. The TCL1 positive control was a stable MSCV-TCL1 retrovirally transfected 2B4 T cell line (13).
Figure S5. Aneuploidy and chromosome translocations in TCL1-tg BLL (PDF, 78.4 KB) -
SKY analysis of a metaphase chromosome spread from a BLL (K10) (1). Yellow box, Ts(15); red boxes, reciprocal t(2;5); white box, t(X;7).
Figure S6. Aicda expression is elevated in TCL1-tg B cell lymphomas (PDF, 11.8 KB) -
Results are shown as Aicda expression normalized to the expression of a 36B4 control sequence in each sample using QPCR (Supplement Ref. 5). Purified WT cells or whole spleen cells from WT and TCL1-tg mice were tested, along with a panel of 7 different TCL1-tg tumors. TCL1-tg tumors 1 and 6 are DLBCL-centroblast type and tumors 2 — 5 are BLL. K7 is a previously described BLL (12) and K7* is harvested ascites from re-established K7 tumor cells injected into syngeneic mice.