Supplement Text File 2. Cytogenetic and FISH analysis of cell lines
Cell lines were processed for cytogenetic analysis using standard chromosome
preparation methods and GTG banding. More than 20 cells were completely analyzed
for each cell line.
Interphase FISH analysis was performed using methods recommended by the
manufacturer (Bcr/Abl DF probe set, Abbott Molecular, Inc.). Two hundred interphase
cells were analyzed per cell line.
Karyotypes for each cell line are as follows:
BV-173
48,X,-Y,trp(1)(q32q41),del(6)(q21q25),i(8)(q10),der(9)t(8;9)(q11.2;q12),
t(9;22)(q34;q11.2),del(12)(p12p13),add(16)(q22),+der(22)t(9;22)x2,+mar[20]
BV-173R
48,X,-Y,trp(1)(q32q41),t(5;7)(q11.2;p22),del(6)(q21q25),der(8)t(7;8)(q11.2;p?21),+9,
der(9)t(8;9)(q11.2;q12)x2,t(9;22)(q34;q11.2),del(12)(p12p13),add(16)(q22),
+der(22)t(9;22)x2[20]
Three related cells with nonclonal abnormalities were also found among the BV-173
cells; one nonclonal cell was found for BV-173R.
Each cell line had several divergent clonal abnormalities:
BV-173 has normal chromosome 5 and 7 homologues, whereas BV-173R has a
t(5;7)(q11.2;p22), an apparently balanced translocation, in all cells.
BV-173 has an i(8)(q10), whereas BV-173R has a der(8)t(7;8)(q11.2;p?21), which
results in loss of one copy of 8q and gain of another copy of 7q (relative to BV-173).
BV-173 has one der(9)t(8;9)(q11.2;q12), whereas BV-173R has two copies of the
der(9)t(8;9), which results in gains of another copy of 9p and 8q.
BV-173 has a small marker chromosome (+mar), whereas BV-173R has no marker.
FISH Results:
The two cell lines had very similar results from the BCR/ABL FISH analysis: three
to four BCR/ABL fusion signals per cell line were present in each cell. For BV-173, 17%
had three fusion signals and 83% had four fusion signals. This is consistent with the
gains of one or two Philadelphia chromosomes [der(22)t(9;22)] noted by cytogenetic
analysis. BV-173R had 13.5% with three fusion signals [+der(22)t(9;22)] and 86.5% had
four fusion signals [+der(22)t(9;22)x2]. The normal ABL signal was missing in every cell
in both lines. This is also consistent with the standard cytogenetic analysis, which
showed one der(9)t(9;22) and no normal copies of 9q in each cell.