Supplementary Figure 1 - Download as DOC

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Supplementary Figure 1. Flow cytometric analysis of peripheral blood leukocytes
from a healthy control.
CD45-positive peripheral blood lymphocytes from a healthy volunteer were
double-stained with either CD25 and CD4 (a) or CD4 and CADM1 (b). The
percentages of each cell fraction are shown in the figures.

Supplementary Figure 2. Immunomagnetic separation for enrichment of
CD4+CADM1+ cells from the peripheral blood of ATLL patients.
Mononuclear cell fractions (MNC) from four patients with various kinds of ATLL were
separated by magnetic beads with an anti-CADM1 antibody. The left columns show the
FCM analysis of the MNC stained with CD4 and CADM1 before separation. The
middle columns show the FCM analysis of the negative CADM1 fraction after the
separation of the MNC of the patients. The right columns show the FCM analysis of the
positive CADM1 fractions after the separation of the MNC after the separation. The
percentages of cells are shown in each FCM figure at the right upper corner. N.D.; not
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Supplementary Figure 3. Correlation between the percentages of CD4+CADM1+
cells and the percentages of abnormal lymphocytes in PBMCs of HTLV-1 carriers.
The percentage of the CD4+CADM1+ fraction was compared with the percentage of
abnormal lymphocytes in HTLV-1 carriers.

Supplementary Figure 4. Relationship between the serum levels of soluble CADM1
and the percentage of peripheral blood CD4+CADM1+ cells in ATLL patients.
The percentages of CD4+CADM1+ cells in PBMCs were compared with the serum
levels of soluble CADM1 in 2 acute-type, 2 chronic-type, 8 smoldering-type and 2
lymphoma-type of ATLL and 2 HTLV-1 carriers. The intensity of the immunoreactive
band of soluble CADM1 was quantified and expressed as in Figure 4b. The data are
represented by dot plot (a) and box plot (b) graphs. In a, the data from ATL cases and
HTLV-1 carriers are indicated by black circles, and the data from ATL lymphoma cases
are indicated by white circles. In b, a star indicates a significant difference between the
band intensities of the groups (*P<0.01).

Supplementary Figure 5. No expression of CADM1 in non-ATLL lymphomas.
(a) Immunostaining of CADM1 in B-cell acute lymphoblastic leukemia (B-ALL) cell
lines (BALM1 [a1] and BALL1 [a2]) and T-ALL cell line MOLT4 (a3) using an
anti-CADM1 monoclonal (1-10C) antibody. Scale bar, 20 μm. (b) Immunoblot analysis   Formatted: Font color: Auto

in various leukemia cell lines with a chicken anti-human CADM1 antibody (3E1).
-tubulin was used as a loading control.

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