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Figure S4. Overexpression of MYC in pre-cancerous thymocytes does not lead to repression
of genes methylated in lymphomas
(A) Real-time RT-PCR analysis of MYC and nucleolin (nuc) expression in unsorted (gray bars),
EGFP-positive (black bars) and EGFP-negative (white bars) cell populations sorted from
thymocytes of 31 days old EμSR-tTA;Teto-MYC;Teto-Cre;Rosa26LOXPEGFP mouse as described
in Figure 2B. Expression data are presented as fold-increase relative to the EGFP-negative
population. Representative example from three independent experiments is shown.
(B) Expression of genes near the RLGS fragments as determined by Real-time RT-PCR analysis
of EGFP-negative (white bars) and EGFP-positive (black bars) cell populations sorted from
thymocytes of 31 days old EμSR-tTA;Teto-MYC;Teto-Cre;Rosa26LOXPEGFP mouse as described
in S4A. Expression of each gene in EGFP-positive population is presented as fold-decrease
relative to the expression of the same gene in EGFP-negative population. Representative
example from three independent experiments is shown.
(C) FACS profiles on unsorted, GFP-negative and GFP-positive thymocytes isolated from 42
days old EμSR-tTA;Teto-MYC;Teto-Cre;Rosa26LOXPEGFP mouse showing almost identical
expression of CD4 and CD8 markers. The percentage of cells that are in each quadrant of the
FACS profile is indicated.
(D) Real-time RT-PCR analysis of MYC and nucleolin (nuc) expression in thymocytes isolated
from 42 days old EμSR-tTA;Teto-MYC;Teto-Cre;Rosa26LOXPEGFP mouse. Analysis was
perfomed as in A part of this Figure.
(E) The same set of genes as in B part of this Figure was analyzed in thymocytes isolated from
from 42 days old EμSR-tTA;Teto-MYC;Teto-Cre;Rosa26LOXPEGFP mouse.
Figure S4
A
B
MYC
15.00
15
3
Fold reduction in expression
5
10
10.00
4
3
5
5.00
2
1
0
-
+
Unsort.
0
0.00
31 day-old
nuc
7
6
Unsort.
Fold induction in gene expression
31 day-old
EGFP
sorted
C
-
2
1
0
+
EGFP
sorted
Gene loci associated with RLGS fragment
EGFP negative
unsorted
EGFP positive
77
5
77
5
76
2
16
2
16
2
17
CD8
5
CD4
D
E
MYC
nuc
7
6
25
5
20
4
15
3
10
2
5
1
EGFP
sorted
+
0
Unsort.
0
42 day-old
Fold reduction in gene expression
30
Unsort.
Fold induction in gene expression
42 day-old
EGFP
sorted
+
3
2
1
0
Gene loci associated with RLGS fragment
Figure S4