Immunohistochemical analysis of vimentin expression in myocardial tissue from autopsy cases of ischemic heart disease

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Figure legends

Figure 1 Basic vimentin staining pattern in myocardial tissue.

Myocardial cells were negative for vimentin (A); non-myocardial cells, including vascular

endothelium, vascular smooth muscle (S), fibroblasts, nerve fibers (N), adipocytes, and

mesothelial cells (not shown) showed positivity (B). (AB: magnification x100)

Figure 2 Vimentin expression in human myocardial tissue with ischemic heart disease

(positivity in remodeling foci)

Hematoxylin and eosin staining (A, C, E) and vimentin immunohistochemistry (B, D, F) were

used. A, B: Vimentin expression in necrotic myocardium; necrotic myocardium was negative

for vimentin. C, D: Vimentin expression in ongoing remodeling of the myocardium;

upregulated vimentin expression was observed around myocardial cells undergoing

remodeling. E, F: Vimentin expression in completely fibrotic foci was not upregulated. (A-F:

magnification x100)

Figure 3 Vimentin expression in human myocardial tissue with ischemic heart disease

(positivity in inflammatory foci)

Results of hematoxylin and eosin staining (A, D), vimentin immunohistochemistry (C, F),

CD68 (B) and myeloperoxidase (MPO) immunohistochemistry (E). A–C: In inflammatory

foci of post-myocardial infarction epicardium, macrophages and fibroblasts that appeared

within the area were positive for vimentin. D–F: Neutrophils infiltrating necrotic myocardium

were positive for vimentin. (ABC: magnification x40, DEF: magnification x100)

Figure 4 Vimentin expression in a mouse model of acute myocardial infarction (MI)

Hematoxylin and eosin staining (A, D, G, J), Azan staining (B, E, H, K), and vimentin

immunohistochemistry (C, F, I, L) were used to examine myocardial tissues. A–C: Normal

tissue, with vimentin expression in vascular endothelial cells between myocardial cells,

similar to human samples. D–F: 3 hours after inducing acute MI, there was no significant

change in vimentin expression. G–I: 1 day after inducing acute MI, there was no fibrosis;

infiltration of inflammatory cells are highlighted (arrows). J–L: 1 week after inducing acute

MI, there was elevated vimentin expression in juvenile fibrous tissue growth with

vascularization. (A-L: magnification x100)

Table 1

Clinical characteristics of the 26 study patients with ischemic heart disease.

including age, sex, diagnosis at autopsy, relative coronary atherosclerosis, whether

resuscitation was performed, whether myocardial necrosis could be observed, and estimated

age of myocardial necrosis, as well as identification of cases used for the figures.

m, male; f, female; AMI, acute myocardial infarction; IHD, ischemic heart disease

Supplementary Figure 1

Tissue samples of human myocardium with ischemic heart disease stained with

phosphotungstic acid hematoxylin (A), vimentin immunohistochemistry (B, D), and

hematoxylin and eosin (C). A, B: Contraction band necrosis was not visualized by

immunostaining for vimentin. C, D: Myocardial congestion was highlighted by

immunostaining for vimentin. (AB: magnification x100, CD: magnification x40)

Supplementary Figure 2

Double immunofluorescence staining for vimentin and CD31. Human myocardial tissue

with fibroblastic and endothelial proliferation (A: hematoxylin and eosin staining)

subjected to immunofluorescence staining with monoclonal antibodies against CD31

(B: green) and vimentin (C: red). Nuclei were stained with DAPI. Some of the

vimentin-positive cells were also CD31-positive endothelial cells (D: merging of B and

C). (A-D: magnification x100)

100m

100m

Fig. 1

100m

100m

100m

100m

100m

Fig. 2

100m

200m

100m

CD68

MPO

200m

100m

vimentin

vimentin

200m

Fig. 3

100m

Vimentin

Azan

control

100m

100m

100m

100m

100m

100m

100m

100m

100m

100m

100m

100m

3 hours

1 day

7 days

Fig. 4

100m

200m

100m

200m

Supplementary Figure 1

HE

CD31

100m

vimentin

merge

Supplementary Figure 2

Case

age

sex

diagnosis at autopsy

coronary atherosclerosis resuscitation myocardial necrosis, age of necrosis

63

AMI, cardiac tamponade

moderate

48

IHD

moderate

68

IHD

severe

54

AMI, recurrent

severe

51

AMI, recurrent

mild

63

IHD

severe

78

AMI, cardiac tamponade

severe

49

IHD

severe

36

IHD

mild

10

47

IHD

severe

11

75

IHD

mild

12

75

AMI, cardiac tamponade

mild

13

67

IHD

moderate

14

75

IHD

moderate

15

56

IHD

mild

16

85

IHD

mild

17

54

IHD

moderate

18

55

AMI, cardiac tamponade

mild

19

71

IHD

mild

20

54

AMI

severe

21

44

IHD

mild

22

90

IHD

mild

23

54

IHD

mild

24

84

IHD

mild

25

42

IHD

mild

26

82

AMI

mild

figure

○ a few days

Fig. 3D-F

○ several hours

Fig. 2EF

Fig. 2A-D, 3A-C

Supplementary Fig. 1CD

○ a few days

○ a few days

Supplementary Fig. 2

○ several hours

Supplementary Fig. 1AB

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