重複驗證實驗之原始資料

Cancer Res 2013; 73: 953-66 Chromosome Instability modulated by BMI1-AURKA signaling drives Progression of head and neck cancer 第一次重複實驗整理

1B (right panel) OECM‐1‐ sh‐scr

FaDu‐ CDH

1 2

BMI1

FaDu‐BMI1 EV let‐7i

β ‐Actin

Aurora A

p53

β ‐Actin

SCR OECM1‐sh‐scr Bi1 OECM1‐sh‐BMI1‐1 Bi2 OECM1‐sh‐BMI1‐2

1.5

FaDu –BMI1 FaDu ‐ CDH sh‐scr sh‐AURKA

BMI1

Aurora A

Fold change of protein level

4C

3I

OECM‐1‐ sh‐BMI1

MG132 phospho‐p53 (ser315)

1

+ + +

β ‐Actin 0.5 0

BMI1 BMI1

5D

Aurora A AURKA

5G

FaDu‐ FaDu –BMI1 CDH sh‐scr sh‐AURKA

Snail

FaDu – BMI1 FaDu ‐ CDH sh‐scr sh‐AURKA

SAS‐ sh‐scr BMI1

BMI1

Phospho‐AKT (ser 473)

Aurora A

Total AKT

Phospho‐GSK‐3β (ser9)

Phospho‐GSK3β (ser9)

Total GSK‐3β

Total GSK3β

BMI1 Aurora A

S2C

β ‐Actin

β ‐catenin

β‐Actin

Aurora A

S3E OECM‐1‐ sh‐scr

OECM‐1‐ sh‐AURKA 1 2

S3F OECM‐1‐ sh‐scr

BMI1

Aurora A

p53

β ‐actin

MG132 phospho‐p53 (ser315) β ‐actin

+ + +

MG132 phospho‐p53 (ser315) β ‐actin

OECM‐1‐ sh‐BMI1 1 2

+ + +

β ‐actin

SAS‐ sh‐BMI1 1 2

第一次重複實驗原始資料

Fig. 1B BMI1

Aurora A

Fig. 3I BMI1

Fig. 4C BMI1

Fig. 5D BMI1

Aurora A Aurora A

Aurora A

Snail β ‐Actin β ‐Actin

p53

β ‐Actin β ‐Actin

phospho‐p53 (ser315)

β ‐Actin

第一次重複實驗原始資料

Fig. 5G BMI1

Aurora A

Phospho‐GSK‐3β (ser9)

Fig. S2C BMI1

Fig. S3E

Fig. S3F

BMI1

Aurora A

Phospho‐AKT (ser 473)

Total AKT

Phospho‐GSK3β (ser9)

p53

β ‐Actin

phospho‐p53 (ser315)

Total GSK‐3β

β ‐Actin Total GSK3β β ‐Actin

β ‐catenin

Aurora A

β ‐actin

β ‐Actin

第二次重複實驗整理

1B (right panel) OECM‐1‐ sh‐scr

4C

3I

OECM‐1‐ sh‐BMI1 1 2

BMI1

Aurora A

FaDu‐ CDH

β ‐Actin SCR OECM1‐sh‐scr Bi1 OECM1‐sh‐BMI1‐1 Bi2 OECM1‐sh‐BMI1‐2

1.5 Fold change of protein level

FaDu –BMI1 FaDu ‐ CDH sh‐scr sh‐AURKA

FaDu‐BMI1 p53

EV let‐7i

BMI1

β ‐Actin

Aurora A

MG132 phospho‐p53 (ser315)

β ‐Actin

1

+ + +

β ‐Actin

0.5 0

BMI1 1

5D

Aurora A 2

5G

FaDu‐ FaDu –BMI1 CDH sh‐scr sh‐AURKA

Snail

FaDu – BMI1 FaDu ‐ CDH sh‐scr sh‐AURKA

SAS‐ sh‐scr BMI1

BMI1

Phospho‐AKT (ser 473)

Aurora A

Total AKT

Phospho‐GSK‐3β (ser9)

Phospho‐GSK3β (ser9)

Total GSK‐3β

Total GSK3β

BMI1 Aurora A

S2C

β ‐Actin

β ‐catenin

β‐Actin

Aurora A

S3E OECM‐1‐ sh‐scr

OECM‐1‐ sh‐AURKA 1 2

S3F OECM‐1‐ sh‐scr

BMI1

Aurora A

p53

p53 β ‐actin

β ‐actin MG132 phospho‐p53 (ser315) β ‐actin

OECM‐1‐ sh‐BMI1 1 2

+ + +

MG132 phospho‐p53 (ser315) β ‐actin

+ + +

β ‐actin

SAS‐ sh‐BMI1 1 2

第二次重複實驗原始資料

Fig. 1B BMI1

Fig. 3I

Fig. 4C

BMI1

Fig. 5D BMI1

Aurora A Aurora A

Snail β ‐Actin

p53 β ‐Actin β ‐Actin β ‐Actin

phospho‐p53 (ser315)

β ‐Actin

第二次重複實驗原始資料

Fig. 5G BMI1

Fig. S2C BMI1

Aurora A

Fig. S3E

Fig. S3F

BMI1

Aurora A

Phospho‐AKT (ser 473)

p53

Phospho‐GSK‐3β (ser9) Total AKT β ‐Actin

Total GSK‐3β

Phospho‐GSK3β (ser9) phospho‐p53 (ser315)

β ‐Actin

Total GSK3β

β ‐catenin

Aurora A

β ‐actin

β ‐Actin

phospho‐p53 (ser315)

β ‐Actin

第三次重複實驗整理

1B (right panel) OECM‐1‐ sh‐scr

4C

3I

OECM‐1‐ sh‐BMI1 1 2

BMI1

Aurora A

FaDu‐ CDH

β ‐Actin SCR OECM1‐sh‐scr Bi1 OECM1‐sh‐BMI1‐1 OECM1‐sh‐BMI1‐2 Bi2

1.5 Fold change of protein level

FaDu –BMI1 FaDu ‐ CDH sh‐scr sh‐AURKA

FaDu‐BMI1 p53

EV let‐7i

BMI1

β ‐Actin

Aurora A

MG132 phospho‐p53 (ser315)

β ‐Actin

1

+ + +

β ‐Actin 0.5 0 BMI1 1

5D

Aurora A 2

5G

FaDu‐ FaDu –BMI1 CDH sh‐scr sh‐AURKA

Snail

FaDu – BMI1 FaDu ‐ CDH sh‐scr sh‐AURKA

SAS‐ sh‐scr BMI1

BMI1

Phospho‐AKT (ser 473)

Aurora A

Total AKT

Phospho‐GSK‐3β (ser9)

Phospho‐GSK3β (ser9)

Total GSK‐3β

Total GSK3β

BMI1 Aurora A

S2C

β ‐Actin

β ‐catenin

β‐Actin

Aurora A

S3E OECM‐1‐ sh‐scr

OECM‐1‐ sh‐AURKA 1 2

S3F OECM‐1‐ sh‐scr

BMI1

Aurora A

p53

p53 β ‐actin

β ‐actin MG132 phospho‐p53 (ser315) β ‐actin

OECM‐1‐ sh‐BMI1 1 2

+ + +

MG132 phospho‐p53 (ser315) β ‐actin

+ + +

β ‐actin

SAS‐ sh‐BMI1 1 2

第三次重複實驗原始資料

Fig. 1B BMI1

Fig. 3I BMI1

Fig. 4C BMI1

Fig. 5D BMI1

Aurora A

Aurora A Aurora A Snail

β ‐Actin

p53

β ‐Actin β ‐Actin

phospho‐p53 (ser315)

β ‐Actin

第三次重複實驗原始資料

Fig. 5G BMI1

Fig. S2C BMI1

Aurora A

Fig. S3E

Fig. S3F

BMI1

Aurora A

Phospho‐AKT (ser 473)

p53 p53

Phospho‐GSK‐3β (ser9) Total AKT

β ‐Actin β ‐Actin Total GSK‐3β Phospho‐GSK3β (ser9)

phospho‐p53 (ser315) phospho‐p53 (ser315)

β ‐Actin

Total GSK3β

β ‐catenin

Aurora A

β ‐actin

β ‐Actin

Clin Cancer Res 2010; 16: 4561-71 Regulation of excision repair cross-complementation group 1 by Snail contributes to cisplatin resistance in head and neck cancer 第一次重複實驗整理

FADU‐CMV #1 #2 NBS1

FADU‐NBS1 FADU‐NBS1 scr shsnail #1 #2 #1 #2

FADU‐NBS1 #1 #2 95

NBS1 Snail

Snail ERCC1 actin

ERCC1 actin

Cancer Res 2013; 73:4147-57 Connective tissue growth factor activates pluripotency genes and mesenchymal-epithelial transition in head and neck cancer cells

重複實驗整理

Figure S3C: sphere formation in FaDu-Neo vs. FaDuC1/FaDu-C2 100 80 Sphere counts/ 10000 cells

FaDu‐ FaDu‐C1 FaDu‐C2 neo n1 61 92 86 n2 79 85 91 AVE 70 88.5 88.5 STDEV 12.72792 4.949747 3.535534

60 40 20 0 FaDu‐neo FaDu‐C1 FaDu‐C2

Figure S4C: sphere formation in FaDu + citric buffer vs. FaDu + rCTGF 150

AVE

63.333333

STDE V

3.3291641

Fadu+rCTGF 113.5 103.5 122.5 113.166666 7 9.50438495 3

120 Sphere counts/ 10000 cells

N1 N2 N3

Fadu+citric 65.5 65 59.5

90 60 30 0 Fadu+citric Fadu+rCTGF

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