Supplemental Table 2 RPPA results of the murine data set
ischemia RT [h]
0
p-HER2 (Y1248)
43
26
25
25
26
90
89
81
76
82
83
72
71
75
91
114
79
85
84
108 102
71
85
73
63
73
90
76
69
68
47
62
53
49
32
24
36
74
68
68
51
57
55
49
43
37
43
62
45
28
33
37
44
70
72
72
64
CF
66
77
69
69
SF 100
61
56
72
51
SF
SF
SF
CF
p-Akt (S473)
100
100
SF
100
100
CF
p-GSK3beta (S9)
SF
100
CF
p-PRAS40 (T246)
SF
100
CF
p-mTOR (S2448)
p-4E-BP1 (T37/46)
5
54
100
CF
p-PTEN (S380)
3
47
CF
p-EGFR (Y1148)
1
45
SF
CF
p-HER3 (Y1289)
0.5
SF
100
ischemia 4°C [h]
24
17
36
73
30
34
29
20
35
0
0.5
1
3
5
Sensitivity
24
73
98
49
57
30
68
76
52
57
17
113 100
89
101
75
90
88
82
80
62
78
75
83
80
95
74
100
75
68
56
81
86
89
97
54
73
91
67
81
52
74
76
52
36
41
67
70
51
48
26
85
76
82
80
53
66
73
73
68
36
51
69
75
47
30
50
72
64
46
25
80
77
86
80
64
77
85
75
75
56
100 116 117 127
93
63
100
100
100
100
100
100
100
100
against
ischemia
Relative signal intensity
difference between SF vs.
CF reference (CF set as
100%)
+++
59
+
88
+
174
+
98
+++
81
++
73
++
86
+
105
++
84
49
CF
p-S6RP (S235/236)
SF
100
CF
p-p38 MAPK (T180/Y182) SF
CF
p-Erk-1/2 (T202/Y204)
SF
CF
p-NFkappaB p65 (S536)
SF
100
100
100
CF
p-STAT3 (S727)
SF
CF
p-STAT5 (Y694)
SF
CF
HER2 DAKO
SF
100
100
100
CF
HER2 CB11
SF
CF
ERalpha
SF
CF
PgR
SF
CF
100
100
100
45
61
42
109 102
85
92
100 111
91
101
100
78
83
69
65
62
54
43
33
28
34
25
22
22
23
21
50
57
64
49
30
34
37
34
71
97
100
76
70
95
94
89
87
94
111
96
94
115 130
89
78
94
86
87
85
104
91
104
83
85
86
85
87
96
86
98
74
76
80
74
97
113
91
89
40
83
91
57
44
74
88
96
52
60
30
16
24
50
100
49
64
50
40
108
100
100
100
100
100
100
100
100
100
100
75
102
94
41
114 110 120
99
98
97
81
86
107
94
108
97
111 112
70
80
97
86
99
42
41
48
51
29
19
35
62
39
26
15
65
89
65
69
39
63
70
51
62
26
82
68
84
94
84
71
87
74
90
78
113 138 111 112
91
137
94
102 128
87
84
88
74
101
61
87
83
79
87
52
76
87
80
82
68
90
90
81
88
69
82
60
85
76
78
92
109
85
89
88
42
45
64
77
44
49
82
55
69
47
-
90
++
65
+++
78
++
65
+
100
-
109
+
114
-*
111
+
119
++
114
Akt
SF
CF
mTOR
SF
100
100
CF
Cyclin D1
SF
CF
Cytokeratin 18
SF
CF
GAPDH
SF
CF
beta-Actin
SF
CF
100
100
100
100
97
110 106 101
93
119 114 109
35
69
84
56
42
72
88
93
82
113 119
92
71
107 117 101
129 113
82
77
62
111
78
85
110 143
85
98
85
107
96
88
92
95
92
95
108 129 107
91
61
43
85
28
77
64
100
100
100
100
100
100
110
98
151 130 117
104
80
126 125
79
44
41
61
69
42
45
88
56
68
43
84
88
129 119 106
86
83
101 112
89
111
86
105
96
62
65
69
87
81
40
109 119 119 143
77
81
88
81
92
58
109
79
95
98
86
103 105 102
94
87
-
101
++
122
+
99
+
70
-
78
-
120
The results are depicted as the mean normalized signal intensity, relative to the respective reference sample (0 h ischemia) which was set as 100%.
p-Hsp27 (S82) was not analyzable because protein expression was too weak.
= ≥ 25% loss of signal in one or more ischemia
+
samples compared to reference sample (time point 0) which was set as 100%; 24 h excluded
= ≥ 50% loss of signal in one or more ischemia
++
samples compared to reference sample (time point 0) which was set as 100%; 24 h excluded
= ≥ 75% loss of signal in one or more ischemia
+++
samples compared to reference sample (time point 0) which was set as 100%; 24 h excluded
(24 h excluded because after 24 h nearly each protein showed massive loss of signal and the other time points are clinically more
relevant)
*
= not sensitive
=
the antibody against HER2 clone CB11 is a
monoclonal antibody; thus the signals were in general weaker than those revealed by the DAKO
antibody against HER2 which is a polyclonal one
SF
= standard formalin
CF
= cold formalin
RT
= room temperature