Fig S1. Model for Tam action
CoA
?
CoR
CoA
CoR
CoR
ER
Tam
ER
CoR
CoR
ERE
MCF7
+
Tam binding to ER changes
conformation of ER which can not
bind to ERE or Tam bound ER is
complexed
with
co-repressor
which inhibits transcription
Co-Repressors block
Transcription
Tam
MAPK Kinase Signalling
Other signaling pathways
??
Apoptosis and/
or growth arrest
Fig S2. Pie-chart distribution of identified proteins on the
basis of cellular functional and localization
a).
b).
Cell Membrane
Mitochondria
Golgi Complex
Differentiation
RNA processing
Heat shock
proteins Glucose
metabolic
pathway
Structural
ER
Nucleus
Cytoskeleton
Protein
folding
Apoptosis
Cytoplasm
Encirled precursor ions were picked for MS/MS among many others
1939.9130
1556.7327
1804.9077
Fig. S3 a: Peptide mass fingerprint of spot no.3 identified as ENO-1
MS/MS
VVIGMDVAASEFFR
1556.7327 (Ion Score 30)
AAVPSGASTGIYEALELR
LAMQEFMILPVGAANFR
1804.9077 (Ion Score 95)
1939.9130 (Ion Score 31)
Fig. S3 b: Mascot Search Result of spot 3 based on M/S and MS/MS peptides and
ion masses
precursor ions are denoted as underlined
1440.7086
1166.5831
1298.6337
Fig S4 a: Peptide mass fingerprint of spot no. 46 identified as E6AP
picked precursor ions for MS/MS
MS/MS
DHIIDDALVR
1166.5831 (Ion Score 43)
LFLQFTTGTDR
1298.6337 (Ion Score 10)
VFSSAEALVQSFR
1440.7086 (Ion Score 14)
Fig S4 b: Mascot Search Result of spot 46 based on M/S and MS/MS peptides
and ion masses
precursor ions are denoted as underlined
1344.7874
1149.6665
1051.5275
Fig S5 a: Peptide mass fingerprint of spot no. 47 identified as NM23
MS/MS
1051.5275 Ion Score 13
1149.6665 Ion Score 23
GDFCIQVGR
DRPFFAGLYK
1344.7874 Ion Score 25
TFIAIKPDGVQR
Fig S5 b: Mascot Search Result of spot 47 based on M/S and MS/MS peptides and
ion masses
precursor ions are denoted as underlined
Fig S6 : Graphical representation for Densitometry analysis of all western Blots
Relative Expression
a.
b.
1.5
1
0.5
0
MCF7 unind
Tam
Relative Expression
Fig.2
MCF7 Untreated
Tam
1.2
0.8
0.4
0
ENO1
NM23
1.5
b.
1
0.5
0
0h
24h
48h
72h
1.5
Relative Expression
a.
Relative Expression
Fig.3
1
0.5
0
0h
24h
48h
72h
1.2
d.
Relative Expression
Relative Expression
c.
1.2
0.8
0.4
0
unind
5uM
1
0.8
0.6
0.4
0.2
0
7.5uM 10uM
unind
Fig.5
c.
b.
1.5
1
0.5
0
NF CF
7.5uM Tam (h)
0
NF CF
NF CF
NF CF
1
3
6
Relative Expression
Relative Expression
2
2.5
2
1.5
1
0.5
0
5uM
7.5uM
10uM
Fig.7
Relative Expression
d.
a.
50
40
30
20
10
0
Relative Expression
Fig.6
1.5
1
0.5
0
MCF7
MCF7+
siE6AP