Endotoxin Leakage and Toll-like Receptor 4 Up-regulation Promote

Doxorubicin-caused Systemic Inflammation

Lintao Wang1, Qian Chen1, Haixia Qi1, Chunming Wang2, Cheng Wang1,3, Junfeng Zhang1,4,*, Lei Dong1,*

1. State Key Laboratory of Pharmaceutical Biotechnology, NJU Advanced Institute for Life Sciences (NAILS), School of life sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210093, China.

2. State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Taipa, Macau SAR.

3. Department of Clinical Laboratory, Jinling Hospital, Nanjing University School of Medicine, 305 East Zhongshan Road,Nanjing, 210002, China

4. Jiangsu Provincial Laboratory for Nano-Technology, Nanjing University, Nanjing, P. R. China

* Corresponding author at: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin avenue, Nanjing 210093, China.

E-mail addresses: jfzhang@nju.edu.cn (J. Zhang), leidong@nju.edu.cn (L. Dong).

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Catalogue

1. Table S1…………………………………………………………………………...Page 3

2. Supplementary Fig S1……………………………………………………………Page 4

3. Supplementary Fig S2……………………………………………………….…..Page 5

4. Supplementary Fig S3…………………………………………………….……..Page 6

5. Supplementary Fig S4……………………………………………………….…..Page 7

6. Supplementary Fig S5……………………………………………………….…..Page 8

7. Supplementary Fig S6……………………………………………………….…..Page 9

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Table S1: Information of the clinical sample donors.

Sample no. Gender Age Disease Previous Treatment

Health Donor1 Male 45 - -

2 Female 47 - -

3 Female 51 - -

4 Female 58 - -

5 Male 60 - -

6 Male 52 - -

7 Male 47 - -

8 Female 50 - -

9 Female 59 - -

10 Male 62 - -

Cancer Patient Without DOX Treatment1 Male 47 Lymphoma -

2 Male 64 Lymphoma -

3 Male 52 Lymphoma -

4 Female 53 Lymphoma -

5 Male 60 Lymphoma -

6 Female 59 Lymphoma -

7 Female 68 Gastric Cancer -

8 Male 69 Gastric Cancer -

9 Male 57 Gastric Cancer -

10 Male 58 Gastric Cancer -

Cancer Patient With DOX Treatment1 Female 70 Lymphoma Doxorubicin

2 Female 63 Lymphoma Doxorubicin

3 Male 66 Lymphoma Doxorubicin

4 Female 45 Lymphoma Doxorubicin

5 Male 66 Lymphoma Doxorubicin

6 Male 61 Lymphoma Doxorubicin

7 Female 47 Breast Cancer Doxorubicin

8 Female 65 Breast Cancer Doxorubicin

9 Female 45 Breast Cancer Doxorubicin

10 Female 45 Breast Cancer Doxorubicin

11 Female 59 Breast Cancer Doxorubicin

12 Female 43 Gastric Cancer Doxorubicin

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13 Male 55 Gastric Cancer Doxorubicin

14 Male 66 Gastric Cancer Doxorubicin

Supplementary Fig S1

Supplementary Fig S1. The ET concentration in DGM mice receiving DOX (20

mg/kg body weight) and/or LPS. (n=10). *p < 0.05.

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Supplementary Fig S2

Supplementary Fig S2. Serum level of IL-1β in DGM mice receiving DOX (20 mg/kg

body weight) and/or LPS (n=10). *p < 0.05.

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Supplementary Fig S3

Supplementary Fig S3. Body weights of DGM mice receiving DOX (20 mg/kg body

weight) and/or LPS. LPS at 5 mg/kg body weight killed all the animals at the 3 rd day

(n=10).

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Supplementary Fig S4

Supplementary Fig S4. Evaluation of multi-organ damages in DGM+DOX+LPS

group and in DGM+DOX group. (A) Serum levels of CK, LDH, BUN and ALT in

DGM+DOX, DGM+DOX+2mg/kg LPS and DGM+DOX+5mg/kg LPS mice (n=10). *p

< 0.05 versus DGM+DOX/DGM+DOX+2mg/kg LPS or DGM+DOX+5mg/kg LPS. (B)

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Representative histological images of H&E staining of the heart, kidney and liver of

the experimental mice.

Supplementary Fig S5

Supplementary Fig S5. The influence of Rapamycin on the toxicity of DOX, as

evidenced by the change in body weight (left panel) and the survival rate (right panel)

of mice receiving DOX (20 mg/kg body weight). (n=10).

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Supplementary Fig S6

Supplementary Fig S6. Multi-organ damages in DOX+Rapamycin group and DOX

group. (A) Serum levels of CK, LDH, BUN and ALT in DOX and DOX+Rapamycin

groups (n = 10). *p < 0.05. (B) Representative histological images of H&E staining of

the heart, kidney, and liver.

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