Newcastle disease virus enhances the growth-inhibiting and - - PowerPoint PPT Presentation

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Newcastle disease virus enhances the growth-inhibiting and - - PowerPoint PPT Presentation

Newcastle disease virus enhances the growth-inhibiting and proapoptotic actions of temozolomide on glioblastoma cells in vitro and in vivo Yang Bai 1 , Yong Chen 1 , Xinyu Hong 1 , Xinrui Liu 1 , Xing Su 2 , Shanji Li 1 , Xuechao Dong 1 , Gang Zhao


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SLIDE 1

Newcastle disease virus enhances the growth-inhibiting and proapoptotic actions of temozolomide on glioblastoma cells in vitro and in vivo

Yang Bai1, Yong Chen1, Xinyu Hong1, Xinrui Liu1, Xing Su2, Shanji Li1, Xuechao Dong1, Gang Zhao1*, Yunqian Li1* 1 Department of Neurosurgery, First Hospital of Jilin University, Changchun, China 2 Laboratory of Cancer Precision Medicine, First Hospital of Jilin University, Changchun, China *Correspondence to Yunqian Li1* 13943188080@163.com Gang Zhao1* zhao_gangjlu@aliyun.com

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SLIDE 2

Supplementary Figure 1: NDV MOI 0 0.1 1 10

45kDa 35kDa 25kDa 15kDa 180kDa 140kDa 100kDa 80kDa 60kDa

Anti-cleaved caspase3 Anti-Parp1 Anti-Bad

45kDa 35kDa 25kDa 15kDa

Anti-Bid

NDV MOI 0 0.1 1 10

25kDa 15kDa 35kDa

Anti-cleaved Bid

25kDa 15kDa 35kDa

Anti-Bax Anti-Bcl-2

25kDa 15kDa 35kDa

Anti-Bcl-xL

25kDa 15kDa 35kDa 45kDa

Anti-β-tubulin

80kDa 60kDa 45kDa 35kDa

Supplementary Figure 1: Uncropped western blots used in figure 3 (A). The boxes indicate where the figure was cropped in the main text. Supplementary Figure 2: Uncropped western blots used in figure 3 (B). The boxes indicate where the figure was cropped in the main text.

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SLIDE 3

Supplementary Figure 2: NDV MOI 0 0.1 1 10

180kDa 140kDa 100kDa 80kDa 60kDa

Anti-PI3K Anti-p-PI3K

140kDa 100kDa 80kDa 180kDa 80kDa 60kDa 45kDa 35kDa

Anti-AKT

80kDa 60kDa 45kDa 35kDa

Anti-p-AKT

NDV MOI 0 0.1 1 10 NDV MOI 0 0.1 1 10

180kDa 140kDa 100kDa 80kDa 60kDa

Anti-mTOR

180kDa 140kDa 100kDa 80kDa

Anti-p-mTOR

80kDa 60kDa 35kDa 45kDa

Anti-AMPK

80kDa 60kDa 35kDa 45kDa 25kDa 15kDa 35kDa 10kDa

Anti-4EBP1

25kDa 15kDa 35kDa 10kDa

Anti-p-4EBP1

80kDa 60kDa 140kDa 100kDa 45kDa

Anti-S6K

80kDa 60kDa 140kDa 100kDa

Anti-p-S6K Anti-p-AMPK

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SLIDE 4

Supplementary Figure 3: NDV moi1 0h12h24h36h48h

T98g

0h12h24h36h48h

LN18

25kDa 15kDa

Anti-cleaved Bid

100kDa 80kDa 60kDa

Anti-Parp1

140kDa 25kDa 15kDa

Anti-cleaved Bid

100kDa 80kDa 60kDa

Anti-Parp1

140kDa

Anti-pAKT

80kDa 60kDa 45kDa 35kDa

Anti-pAKT

80kDa 60kDa 45kDa 35kDa 60kDa 45kDa 35kDa

Anti-p-AMPK

80kDa 60kDa 45kDa

Anti-p-AMPK

80kDa 60kDa 45kDa 35kDa

Anti-β-tubulin

80kDa 60kDa 45kDa 35kDa

Anti-β-tubulin

80kDa

Supplementary Figure 3: Uncropped western blots used in figure 3 (C).The boxes indicate where the figure was cropped in the main text.

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SLIDE 5

Supplementary Figure 4: NDV — + — + TMZ — — + + NDV — + — + TMZ — — + + T98G LN18 NDV — + — + TMZ — — + + NDV — + — + TMZ — — + + T98G LN18

45kDa 35kDa 25kDa 15kDa

Anti-cleaved caspase3

45kDa 35kDa 25kDa 15kDa

Anti-cleaved caspase3

180kDa 140kDa 100kDa 80kDa 60kDa 180kDa 140kDa 100kDa 80kDa 60kDa

Anti-Parp1 Anti-Parp1

25kDa 15kDa 35kDa 25kDa 15kDa 35kDa

Anti-cleaved Bid Anti-cleaved Bid

25kDa 15kDa 35kDa 25kDa 15kDa 35kDa

Anti-Bax Anti-Bax

25kDa 15kDa 35kDa 25kDa 15kDa 35kDa 25kDa 15kDa 35kDa 45kDa 25kDa 15kDa 35kDa 45kDa 80kDa 60kDa 45kDa 35kDa 80kDa 60kDa 45kDa 35kDa

Anti-Bcl-2 Anti-Bcl-2 Anti-Bcl-xL Anti-Bcl-xL Anti-β-tubulin Anti-β-tubulin

Supplementary Figure 4: Uncropped western blots used in figure 4 (A).The boxes indicate where the figure was cropped in the main text.

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SLIDE 6

Supplementary Figure 5: NDV — + — + TMZ — — + + NDV — + — + TMZ — — + + T98G LN18 NDV — + — + TMZ — — + + NDV — + — + TMZ — — + + T98G LN18

180kDa 140kDa 100kDa 80kDa 60kDa

Anti-PI3K

180kDa 140kDa 100kDa 80kDa 60kDa

Anti-PI3K

180kDa 140kDa 100kDa 80kDa 60kDa 180kDa 140kDa 100kDa 80kDa 60kDa

Anti-p-PI3K

80kDa 60kDa 45kDa 35kDa 80kDa 60kDa 45kDa 35kDa 60kDa 45kDa 35kDa

Anti-p-AKT

80kDa 60kDa 45kDa 35kDa

Anti-p-AKT

80kDa

Anti-p-PI3K Anti-AMPK Anti-AKT Anti-AKT Anti-AMPK Anti-p-AMPK Anti-p-AMPK

80kDa 60kDa 45kDa 35kDa 80kDa 60kDa 45kDa 35kDa 60kDa 45kDa 35kDa 80kDa 60kDa 45kDa 35kDa 80kDa 180kDa 140kDa 100kDa 80kDa

Anti-mTOR

180kDa 140kDa 100kDa 80kDa

Anti-mTOR

180kDa 140kDa 100kDa 80kDa

Anti-p-mTOR

180kDa 140kDa 100kDa 80kDa

Anti-p-mTOR

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SLIDE 7

Supplementary Figure 6: NDV — + — + TMZ — — + + NDV — + — + TMZ — — + + T98G LN18 NDV — + — + TMZ — — + + NDV — + — + TMZ — — + + T98G LN18

25kDa 15kDa 35kDa 10kDa

Anti-4EBP1

25kDa 15kDa 35kDa 10kDa

Anti-4EBP1

25kDa 15kDa 35kDa 10kDa

Anti-p-4EBP1

25kDa 15kDa 35kDa 10kDa

Anti-p-4EBP1

80kDa 60kDa 140kDa 100kDa 45kDa

Anti-S6K

80kDa 60kDa 140kDa 100kDa 45kDa

Anti-S6K

80kDa 60kDa 100kDa

Anti-p-S6K

80kDa 60kDa 100kDa

Anti-p-S6K

140kDa 140kDa

Supplementary Figure 5-6: Uncropped western blots used in figure 4 (B).The boxes indicate where the figure was cropped in the main text.

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SLIDE 8

Supplementary Figure 7: NDV — + — + TMZ — — + + NDV — + — + TMZ — — + + NDV — + — + TMZ — — + + U251 U87 C6

80kDa 60kDa 45kDa 35kDa

Anti-p-AKT

60kDa 45kDa 35kDa

Anti-p-AKT

80kDa 60kDa 45kDa 35kDa

Anti-p-AKT

80kDa 80kDa 60kDa 45kDa 35kDa

Anti-β-tubulin

80kDa 60kDa 45kDa 35kDa

Anti-β-tubulin

80kDa 60kDa 45kDa 35kDa

Anti-β-tubulin

Supplementary Figure 7: Uncropped western blots used in figure 4 (C).The boxes indicate where the figure was cropped in the main text.

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SLIDE 9

Supplementary Figure 8: T98G LN18 T98G LN18

Compound C NDV moi1 TMZ 200μM

- + + + - + - + - - + + - + + + - + - + - - + + - + + + - + - + - - + + - + + + - + - + - - + +

80kDa 60kDa 45kDa 35kDa 80kDa 60kDa 45kDa 35kDa 80kDa 60kDa 45kDa 35kDa

Anti-AMPK Anti-β-tubulin Anti-p-AMPK

80kDa 60kDa 45kDa 35kDa 80kDa 60kDa 45kDa 35kDa 80kDa 60kDa 45kDa 35kDa

Anti-AMPK Anti-β-tubulin Anti-p-AMPK

MK-2206 NDV moi1 TMZ 200μM

Anti-β-tubulin Anti-β-tubulin

80kDa 60kDa 45kDa 35kDa 80kDa 60kDa 45kDa 35kDa

Anti-p-AKT

60kDa 45kDa 35kDa 80kDa

Anti-p-AKT

60kDa 45kDa 35kDa 80kDa

Anti-AKT

80kDa 60kDa 45kDa 35kDa

Anti-AKT

80kDa 60kDa 45kDa 35kDa

Supplementary Figure 8: Uncropped western blots used in figure 4 (D&F).The boxes indicate where the figure was cropped in the main text.

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SLIDE 10

Supplementary Figure 9:

AT13148 TMZ 200μM

- + - + - - + +

T98g

80kDa 60kDa 45kDa 35kDa

Anti-AKT Anti-p-AKT

60kDa 45kDa 80kDa 25kDa 15kDa

Anti-cleaved Bid

80kDa 60kDa 45kDa 35kDa

Anti-β-tubulin

140kDa 100kDa 80kDa 60kDa

Anti-Parp1

- + - + - - + +

LN18

140kDa 100kDa 80kDa 60kDa

Anti-Parp1

25kDa 15kDa

Anti-cleaved Bid Anti-p-AKT

60kDa 45kDa 80kDa 60kDa 45kDa 35kDa

Anti-AKT

80kDa 60kDa 45kDa 35kDa

Anti-β-tubulin

Supplementary Figure 9: Uncropped western blots used in figure 4 (I).The boxes indicate where the figure was cropped in the main text.