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 1 *, Yunqian Li 1 * 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 Li 1 * 13943188080@163.com Gang Zhao 1 * zhao_gangjlu@aliyun.com

Supplementary Figure 1: NDV MOI 0 0.1 1 10 NDV MOI 0 0.1 1 10 35kDa 25kDa 45kDa Supplementary Figure 1: 35kDa 15kDa 25kDa Uncropped western blots 15kDa used in figure 3 (A). Anti-cleaved Bid 35kDa Anti-cleaved caspase3 The boxes indicate where the 25kDa figure was cropped in the 180kDa 140kDa 15kDa main text. 100kDa Anti-Bax 80kDa 60kDa 35kDa Anti-Parp1 25kDa 45kDa 15kDa 35kDa 25kDa Anti-Bcl-2 Supplementary Figure 2: 15kDa 45kDa Uncropped western blots 35kDa Anti-Bad used in figure 3 (B). 25kDa The boxes indicate where the 15kDa figure was cropped in the Anti-Bcl-xL main text. 80kDa 60kDa Anti-Bid 45kDa 35kDa Anti- β -tubulin

Supplementary Figure 2: NDV MOI 0 0.1 1 10 NDV MOI 0 0.1 1 10 NDV MOI 0 0.1 1 10 35kDa 180kDa 25kDa 180kDa 140kDa 140kDa 100kDa 100kDa 15kDa 80kDa 80kDa 10kDa 60kDa 60kDa Anti-4EBP1 35kDa Anti-PI3K Anti-mTOR 180kDa 25kDa 140kDa 100kDa 180kDa 80kDa 15kDa 140kDa 10kDa Anti-p-PI3K 100kDa 80kDa Anti-p-4EBP1 80kDa Anti-p-mTOR 140kDa 60kDa 100kDa 80kDa 80kDa 45kDa 60kDa 60kDa 35kDa 45kDa Anti-AKT 45kDa 35kDa Anti-S6K 80kDa Anti-AMPK 60kDa 140kDa 80kDa 100kDa 80kDa 45kDa 60kDa 35kDa 60kDa 45kDa Anti-p-AKT 35kDa Anti-p-S6K Anti-p-AMPK

Supplementary Figure 3: T98g LN18 NDV moi1 0h12h24h36h48h 0h12h24h36h48h 140kDa 140kDa Supplementary Figure 3: 100kDa 100kDa 80kDa 80kDa Uncropped western blots used 60kDa Anti-Parp1 60kDa Anti-Parp1 25kDa in figure 3 (C).The boxes 25kDa indicate where the figure was 15kDa 15kDa cropped in the main text. Anti-cleaved Bid Anti-cleaved Bid 80kDa 80kDa 60kDa 60kDa 45kDa 35kDa 45kDa Anti-pAKT 35kDa Anti-pAKT 80kDa 60kDa 80kDa 45kDa 60kDa 35kDa 45kDa Anti-p-AMPK Anti-p-AMPK 80kDa 80kDa 60kDa 60kDa 45kDa 45kDa 35kDa 35kDa Anti- β -tubulin Anti- β -tubulin

Supplementary Figure 4: T98G LN18 T98G LN18 NDV — + — + NDV — + — + NDV — + — + NDV — + — + TMZ — — + + TMZ — — + + TMZ — — + + TMZ — — + + 45kDa 45kDa 35kDa 35kDa 35kDa 35kDa 25kDa 25kDa 25kDa 25kDa 15kDa 15kDa Anti-cleaved caspase3 15kDa 15kDa Anti-cleaved caspase3 180kDa 140kDa 180kDa 100kDa 140kDa Anti-Bcl-2 100kDa Anti-Bcl-2 80kDa 80kDa 60kDa 60kDa 45kDa Anti-Parp1 Anti-Parp1 35kDa 35kDa 45kDa 25kDa 35kDa 35kDa 25kDa 25kDa 15kDa 25kDa 15kDa 15kDa 15kDa Anti-Bcl-xL Anti-cleaved Bid Anti-Bcl-xL Anti-cleaved Bid 80kDa 35kDa 35kDa 80kDa 60kDa 25kDa 60kDa 25kDa 45kDa 35kDa 45kDa 15kDa 35kDa 15kDa Anti-Bax Anti- β -tubulin Anti-Bax 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.

Supplementary Figure 5: T98G LN18 T98G LN18 NDV — + — + NDV — + — + NDV — + — + NDV — + — + TMZ — — + + TMZ — — + + TMZ — — + + TMZ — — + + 180kDa 180kDa 140kDa 140kDa 80kDa 80kDa 100kDa 100kDa 60kDa 80kDa 60kDa 80kDa 60kDa 60kDa 45kDa 45kDa 35kDa 35kDa Anti-AKT Anti-PI3K Anti-AKT Anti-PI3K 180kDa 80kDa 80kDa 140kDa 60kDa 180kDa 60kDa 100kDa 140kDa 80kDa 45kDa 45kDa 60kDa 100kDa 35kDa 80kDa 35kDa Anti-p-PI3K Anti-p-AKT Anti-p-AKT 60kDa Anti-p-PI3K 80kDa 180kDa 180kDa 60kDa 140kDa 140kDa 100kDa 100kDa 80kDa 45kDa 80kDa 80kDa 60kDa 35kDa Anti-mTOR Anti-mTOR 45kDa Anti-AMPK 35kDa Anti-AMPK 180kDa 180kDa 140kDa 80kDa 140kDa 100kDa 100kDa 60kDa 80kDa 80kDa 80kDa 60kDa 45kDa Anti-p-mTOR Anti-p-mTOR 35kDa 45kDa 35kDa Anti-p-AMPK Anti-p-AMPK

Supplementary Figure 6: T98G LN18 T98G LN18 NDV — + — + NDV — + — + NDV — + — + NDV — + — + TMZ — — + + TMZ — — + + TMZ — — + + TMZ — — + + 140kDa 100kDa 140kDa 35kDa 100kDa 80kDa 35kDa 80kDa 25kDa 60kDa 60kDa 25kDa 45kDa 15kDa 45kDa Anti-S6K 10kDa 15kDa Anti-S6K 10kDa 140kDa Anti-4EBP1 Anti-4EBP1 100kDa 80kDa 140kDa 35kDa 35kDa 100kDa 60kDa 25kDa 80kDa 25kDa Anti-p-S6K 60kDa 15kDa 15kDa 10kDa 10kDa Anti-p-4EBP1 Anti-p-4EBP1 Anti-p-S6K Supplementary Figure 5-6: Uncropped western blots used in figure 4 (B).The boxes indicate where the figure was cropped in the main text.

Supplementary Figure 7: U251 U87 C6 NDV — + — + NDV — + — + NDV — + — + TMZ — — + + TMZ — — + + TMZ — — + + 80kDa 80kDa 80kDa 60kDa 60kDa 60kDa 45kDa 45kDa 45kDa 35kDa 35kDa 35kDa Anti-p-AKT Anti-p-AKT Anti-p-AKT 80kDa 80kDa 60kDa 60kDa 80kDa 60kDa 45kDa 45kDa 35kDa 35kDa 45kDa Anti- β -tubulin 35kDa Anti- β -tubulin 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.

Supplementary Figure 8: LN18 T98G LN18 T98G Compound C － ＋ ＋ ＋ － ＋ ＋ ＋ － ＋ ＋ ＋ － ＋ ＋ ＋ MK-2206 NDV moi1 NDV moi1 － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ － ＋ TMZ 200μM TMZ 200μM － － ＋ ＋ － － ＋ ＋ － － ＋ ＋ － － ＋ ＋ 80kDa 80kDa 80kDa 80kDa 60kDa 60kDa 60kDa 60kDa 45kDa 45kDa 45kDa 45kDa 35kDa 35kDa 35kDa 35kDa Anti-p-AMPK Anti-p-AKT Anti-p-AMPK Anti-p-AKT 80kDa 80kDa 80kDa 80kDa 60kDa 60kDa 60kDa 60kDa 45kDa 45kDa 45kDa 45kDa 35kDa 35kDa 35kDa 35kDa Anti-AMPK Anti-AMPK Anti-AKT Anti-AKT 80kDa 80kDa 80kDa 80kDa 60kDa 60kDa 60kDa 60kDa 45kDa 45kDa 45kDa 45kDa 35kDa 35kDa 35kDa 35kDa Anti- β -tubulin Anti- β -tubulin Anti- β -tubulin Anti- β -tubulin Supplementary Figure 8: Uncropped western blots used in figure 4 (D&F).The boxes indicate where the figure was cropped in the main text.

Supplementary Figure 9: T98g LN18 － ＋ － ＋ － ＋ － ＋ AT13148 TMZ 200μM － － ＋ ＋ － － ＋ ＋ Supplementary Figure 9: 140kDa 140kDa 100kDa 100kDa Uncropped western blots used 80kDa 80kDa 60kDa 60kDa in figure 4 (I).The boxes indicate where the figure was Anti-Parp1 Anti-Parp1 cropped in the main text. 25kDa 25kDa 15kDa 15kDa Anti-cleaved Bid Anti-cleaved Bid 80kDa 80kDa 60kDa 60kDa 45kDa 45kDa Anti-p-AKT Anti-p-AKT 80kDa 60kDa 60kDa 45kDa 35kDa 45kDa Anti-AKT 35kDa Anti-AKT 80kDa 60kDa 80kDa 60kDa 45kDa 35kDa 45kDa Anti- β -tubulin 35kDa Anti- β -tubulin