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Cancer Res 2013; 73: 953-66 Chromosome Instability modulated by BMI1-AURKA signaling drives Progression of head and neck cancer 1. Figure 5I中total GSK-3beta band與Aurora A 類似 答覆: Full blot 顯示並非相同 Total GSK3b

Aurora A

2. Figure 5D中Aurora A與Figure 5G中Aurora A類似, 但兩者beta-actin不同 答覆: Full blot 顯示並非相同 Figure 5D Aurora A

Figure 5G Aurora A

3. Figure 5D中Aurora A與Figure 3I中Aurora A類似 答覆: 原始資料Full blot 顯示並非相同 Figure 3I Aurora A Figure 5D Aurora A


4. Figure 5D中beta-actin 銳化後與Figure 4C由上而下第一組beta-actin類似 答覆: 原始資料Full blot 顯示並非相同 Figure 4C beta-actin (upper)

Figure 5D beta-actin

5. Figure 5D中beta-actin水平翻轉後 與Figure 4C由上而下第二組beta-actin類似 答覆: 原始資料Full blot 顯示並非相同 Figure 5D beta-actin 水平翻轉

Figure 4C beta-actin

6. Figure S3E上下兩組beta-actin相似 答覆: 原始資料Full blot 顯示並非相同,但因圖形過度類似最後發生錯置 Figure S3E beta-actin (上)

Figure S3E beta-actin (下)


7. Figure S2C beta-actin 水平翻轉後與FigureS3E beta-actin類似 答覆: 原始資料Full blot 顯示並非相同 Figure S2C beta-actin 水平翻轉

Figure S3E beta-actin (下)

8. Figure S3F 上下兩組beta-actin 類似 答覆: 原始資料Full blot 顯示並非相同 Figure S3F beta-actin (上)

Figure S3F beta-actin (下)

9. Figure S3F (BMI1) bands與Figure S4A (BMI1)bands銳化後類似 10. Figure S3F (Aurora A) bands與Figure S4A (Aurora A)bands類似 答覆: Figure S3F, Figure S4A為同一細胞,均為OECM1 knockdown AURKA 2 clones vs. control clone, 因此為同一實驗顯示Aurora A knockdown效率,應於Figure S4A BMI1, Aurora A下加上Figure S3F之actin, 顯示與Figure S3F為同一細胞,但與Figure S4A下半 部為不同gel之結果,但本論文中疏失遺漏未加actin, 將進行更正。


11. Figure S3F (BMI1) bands及(Aurora A)bands 與Figure S6A (BMI1)bands及(Aurora A)類 似 答覆: Figure S3F, Figure S6A為同一細胞,均為OECM1 knockdown AURKA 2 clones vs. control clone, 因此為同一實驗顯示Aurora A knockdown效率,應於Figure S6A BMI1, Aurora A下加上Figure S3F之actin, 顯示與Figure S3F為同一細胞,但與Figure S6A下半 部為不同gel之結果,但本論文中疏失遺漏未加actin, 將進行更正。 12. Figure 1B右列beta-actin 與Figure 2A beta-actin bands 類似 答覆: 原始資料Full blot 顯示並非相同,但因圖形過度類似可能最後發生錯置

Figure 1B 右列beta-actin

Figure 2A beta-actin


第一次重複實驗整理 為確認結果之再現性以及我們發現的正確性,本論文在12/17受pubpeer質疑後即開始 三次獨立重複實驗,無論是否有被誤解均一律重作,已於2/17完成三重複實驗,完全驗 證先前之發現,茲將三次重複獨立實驗之整理結果以及原始資料呈現如下。

1B (right panel) OECM‐1‐ sh‐scr

FaDu‐ CDH

1            2

BMI1

FaDu‐BMI1 EV     let‐7i

β ‐Actin

Aurora A

Aurora A

p53

β ‐Actin

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

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

BMI1

Aurora A

Aurora A

p53

p53

β ‐actin

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

BMI1

Aurora A

Aurora A

Phospho‐AKT (ser 473)

Total AKT

Phospho‐GSK3β (ser9)

p53

p53

β ‐Actin

β ‐Actin

phospho‐p53 (ser315)

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

BMI1

Aurora A

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

BMI1

Aurora A

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

BMI1

Fig. 5D BMI1

Aurora A Aurora A

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

BMI1

Aurora A

Aurora A

Phospho‐AKT (ser 473)

p53

p53

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

Total GSK‐3β

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

β ‐Actin

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

BMI1

Aurora A

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

BMI1

Aurora A

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 Aurora A Snail

β ‐Actin

p53

β ‐Actin β ‐Actin

phospho‐p53 (ser315)

β ‐Actin

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

BMI1

Aurora A

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

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 1. Fig. S3B右列(Snail)(前三個)水平翻轉後與左列(ERCC1)bands相似 答覆: 附上投稿時之放大原圖可見顯著不同。刊登版本因解像度減低造成影像類似。 右側Snail

水平翻轉

上為放大後右側Snail,下為ERCC1,放大倍率後即可見顯著不同


雖然放大原圖可見明顯不同,證實為不實指控,本人於12/19在pubpeer遭檢舉後 仍然重新建立stable line,重複實驗驗證本論文之可靠性。此為2017年2月25日完 成之重複實驗之結果。

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 1. Figure 2B(TW2.6-siC1)與Figure S3C(FaDu-Neo)相似 答覆: 經查證原始資料,Figure S3C FaDu-Neo之相片為誤植,並確認僅代表圖形錯誤, 計數並無更動,已去函Cancer Research Editorial Office要求更正 誤植之原圖

更正後

2. Figure S3C(FaDu-C2)與Figure S4C(FaDu-rCTGF)相似 答覆: 經查證原始資料,Figure S4C FaDu-rCTGF之相片為誤植,並確認僅代表圖形錯誤, 計數並無更動,已去函Cancer Research Editorial Office要求更正 誤植之原圖

更正後

3. Figure 5D(Nanog-Luc(mut))與Figure 5F(Sox2-Luc(mut))相似 答覆: 請見後頁放大圖示,以直線測量兩者明顯不同。


Cancer Res 2013; 73:4147-57 (論文序號3) Connective tissue growth factor activates pluripotency genes and mesenchymal-epithelial transition in head and neck cancer cells 茲針對論文序號3之第一及第二項疑義部分,增加補中資料如后。 第三項疑義因論文之圖檔即可知明顯不同,故不另加說明。 1. Figure 2B(TW2.6-siC1)與Figure S3C(FaDu-Neo)相似 答覆: 經查證原始資料,確認Figure S3C FaDu-Neo之相片為誤植,並單純為代表圖形 於最後排版時誤植,計數並無更動。附上Figure 2B以及Figure S3C之原始計數資料以 及留存之相片檔。 Figure 2B TW2.6/sh‐scr TW2.6/sh‐CTGF 201 117 198 102 199.5 109.5 2.121320344 10.60660172

TW2.6-scr

Figure S3C FaDu‐Neo

FaDu‐C1

FaDu‐C2

70 160 168 68 155 176 69 157.5 172 1.414213562 3.535533906 5.656854249

TW2.6-siC1

FaDu-Neo

誤植之 Figure S3C FaDu-Neo

更正後之 Figure S3C FaDu-Neo (紅色框線)

FaDu-C1

FaDu-C2


2. Figure S3C(FaDu-C2)與Figure S4C(FaDu-rCTGF)相似 答覆: 經查證原始資料,確認Figure S4C FaDu-rCTGF之相片為誤植,並單純為代表圖 形於最後排版時誤植,計數並無更動。附上Figure S3C以及Figure S4C之原始計數資料 以及留存之相片檔。 Figure S3C FaDu‐Neo

FaDu‐C1

Figure S4C

FaDu‐C2

70 160 168 68 155 176 69 157.5 172 1.414213562 3.535533906 5.656854249

FaDu-Neo

FaDu-C1

誤植之 Figure S4C FaDu-rCTGF

FaDu-C2

FaDu+citric buffer FaDu+rCTGF 80 188 87 176 83.5 182 4.949747468 8.485281374

FaDuCitric buffer

FaDurCTGF

更正後之 Figure S4C FaDu-rCTGF (紅色框線)

另附上證明,上述檔案為2013/2/26建立,為確切之原始檔。


本論文受質疑之部分,本人於接獲pubpeer質疑後,亦按照下列步驟處 理:(1)於pubpeer上公開說明;(2)對照原始實驗資料,確認何者為非 事實之質疑,何者為不慎誤植;(3)若有誤植處,與期刊聯絡進行勘誤; (4)無論質疑是否為事實,均進行重複實驗以驗證論文之正確性。本部 分之質疑我們也以相同方式處理,目前勘誤已完成,重複實驗亦於 2017年3月25日完成,再次證明本論文實無可質疑之處。重複實驗之 結果顯示如下。

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


Oncogene 2012; 31:2401-11 Connective tissue growth factor modulates oral squamous cell carcinoma Invasion by activating a miR-504-FOXP1 signalling 1. Figure 4a 左列(GAPDH)band 與Figure 4b (GAPDH)band (前二個)類似 答覆: 本論文之此部分為張正琪教授口腔生物所碩士生張致皓所做。經與張老師聯繫, 取得Figure 4a 左側GAPDH為張同學於2010年參加生醫年會之壁報檔,以及Figure 4b 原始圖檔比對,顯示為不同之bands。 張同學於2010年參加生醫年會之壁報檔

Figure 4a GAPDH

Figure 4b GAPDH

由壁報檔之第三條band即可看出兩者為 不同之圖形。即使前兩條bands亦不相同。

回覆科技部(實驗原始資料與重複驗證整理)  
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