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(A) STING transcript levels across TCGA tumor datasets relative to GTEx organ-matched normal tissues. (B) IHC analysis of STING expression in the UCLA PDAC tissue microarray (n = 138). (C) Immunoblot analysis of protein lysates prepared from STING WT and CRISPR-Cas9 KO HS766T or DANG subcutaneous xenograft tumors. (D) IncuCyte live-cell imaging analysis of SUIT2 <t>TetR</t> STINGR284M cells treated +50 ng/mL doxycycline (DOX) ± 500 nM VE-822 (ATR inhibitor [ATRi]) in anchorage-independent cultures (mean ± SD; n = 6; one-way ANOVA corrected for multiple comparisons by Bonferroni adjustment). Images are representative of respective experimental endpoints. (E) Immunoblot analysis of SUIT2 tumors from mice treated ± DOX diet ± AZD-6738 (25 mg/kg b.i.d.) for 3 days. (F) Experimental design to evaluate the impact of ATRi against PDAC tumors with active STING signaling. (G and H) NCG mice were inoculated with subcutaneous (s.c.; G; n = 8) or orthotopic (H; n = 4) SUIT2 TetR STINGR284M tumors. Mice were fed a DOX-supplemented diet starting 7 days after tumor inoculation and treated ±25 mg/kg ATRi AZD-6738 b.i.d. for 26 days (unpaired t test; mean ± SD). Tumor volumes were measured using micro-computed tomography (μCT). Mass of excised tumor tissue is indicated. *p < 0.05; ****p < 0.0001.
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(A) STING transcript levels across TCGA tumor datasets relative to GTEx organ-matched normal tissues. (B) IHC analysis of STING expression in the UCLA PDAC tissue microarray (n = 138). (C) Immunoblot analysis of protein lysates prepared from STING WT and CRISPR-Cas9 KO HS766T or DANG subcutaneous xenograft tumors. (D) IncuCyte live-cell imaging analysis of SUIT2 <t>TetR</t> STINGR284M cells treated +50 ng/mL doxycycline (DOX) ± 500 nM VE-822 (ATR inhibitor [ATRi]) in anchorage-independent cultures (mean ± SD; n = 6; one-way ANOVA corrected for multiple comparisons by Bonferroni adjustment). Images are representative of respective experimental endpoints. (E) Immunoblot analysis of SUIT2 tumors from mice treated ± DOX diet ± AZD-6738 (25 mg/kg b.i.d.) for 3 days. (F) Experimental design to evaluate the impact of ATRi against PDAC tumors with active STING signaling. (G and H) NCG mice were inoculated with subcutaneous (s.c.; G; n = 8) or orthotopic (H; n = 4) SUIT2 TetR STINGR284M tumors. Mice were fed a DOX-supplemented diet starting 7 days after tumor inoculation and treated ±25 mg/kg ATRi AZD-6738 b.i.d. for 26 days (unpaired t test; mean ± SD). Tumor volumes were measured using micro-computed tomography (μCT). Mass of excised tumor tissue is indicated. *p < 0.05; ****p < 0.0001.
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(A) STING transcript levels across TCGA tumor datasets relative to GTEx organ-matched normal tissues. (B) IHC analysis of STING expression in the UCLA PDAC tissue microarray (n = 138). (C) Immunoblot analysis of protein lysates prepared from STING WT and CRISPR-Cas9 KO HS766T or DANG subcutaneous xenograft tumors. (D) IncuCyte live-cell imaging analysis of SUIT2 <t>TetR</t> STINGR284M cells treated +50 ng/mL doxycycline (DOX) ± 500 nM VE-822 (ATR inhibitor [ATRi]) in anchorage-independent cultures (mean ± SD; n = 6; one-way ANOVA corrected for multiple comparisons by Bonferroni adjustment). Images are representative of respective experimental endpoints. (E) Immunoblot analysis of SUIT2 tumors from mice treated ± DOX diet ± AZD-6738 (25 mg/kg b.i.d.) for 3 days. (F) Experimental design to evaluate the impact of ATRi against PDAC tumors with active STING signaling. (G and H) NCG mice were inoculated with subcutaneous (s.c.; G; n = 8) or orthotopic (H; n = 4) SUIT2 TetR STINGR284M tumors. Mice were fed a DOX-supplemented diet starting 7 days after tumor inoculation and treated ±25 mg/kg ATRi AZD-6738 b.i.d. for 26 days (unpaired t test; mean ± SD). Tumor volumes were measured using micro-computed tomography (μCT). Mass of excised tumor tissue is indicated. *p < 0.05; ****p < 0.0001.
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(A) STING transcript levels across TCGA tumor datasets relative to GTEx organ-matched normal tissues. (B) IHC analysis of STING expression in the UCLA PDAC tissue microarray (n = 138). (C) Immunoblot analysis of protein lysates prepared from STING WT and CRISPR-Cas9 KO HS766T or DANG subcutaneous xenograft tumors. (D) IncuCyte live-cell imaging analysis of SUIT2 <t>TetR</t> STINGR284M cells treated +50 ng/mL doxycycline (DOX) ± 500 nM VE-822 (ATR inhibitor [ATRi]) in anchorage-independent cultures (mean ± SD; n = 6; one-way ANOVA corrected for multiple comparisons by Bonferroni adjustment). Images are representative of respective experimental endpoints. (E) Immunoblot analysis of SUIT2 tumors from mice treated ± DOX diet ± AZD-6738 (25 mg/kg b.i.d.) for 3 days. (F) Experimental design to evaluate the impact of ATRi against PDAC tumors with active STING signaling. (G and H) NCG mice were inoculated with subcutaneous (s.c.; G; n = 8) or orthotopic (H; n = 4) SUIT2 TetR STINGR284M tumors. Mice were fed a DOX-supplemented diet starting 7 days after tumor inoculation and treated ±25 mg/kg ATRi AZD-6738 b.i.d. for 26 days (unpaired t test; mean ± SD). Tumor volumes were measured using micro-computed tomography (μCT). Mass of excised tumor tissue is indicated. *p < 0.05; ****p < 0.0001.
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(A) STING transcript levels across TCGA tumor datasets relative to GTEx organ-matched normal tissues. (B) IHC analysis of STING expression in the UCLA PDAC tissue microarray (n = 138). (C) Immunoblot analysis of protein lysates prepared from STING WT and CRISPR-Cas9 KO HS766T or DANG subcutaneous xenograft tumors. (D) IncuCyte live-cell imaging analysis of SUIT2 <t>TetR</t> STINGR284M cells treated +50 ng/mL doxycycline (DOX) ± 500 nM VE-822 (ATR inhibitor [ATRi]) in anchorage-independent cultures (mean ± SD; n = 6; one-way ANOVA corrected for multiple comparisons by Bonferroni adjustment). Images are representative of respective experimental endpoints. (E) Immunoblot analysis of SUIT2 tumors from mice treated ± DOX diet ± AZD-6738 (25 mg/kg b.i.d.) for 3 days. (F) Experimental design to evaluate the impact of ATRi against PDAC tumors with active STING signaling. (G and H) NCG mice were inoculated with subcutaneous (s.c.; G; n = 8) or orthotopic (H; n = 4) SUIT2 TetR STINGR284M tumors. Mice were fed a DOX-supplemented diet starting 7 days after tumor inoculation and treated ±25 mg/kg ATRi AZD-6738 b.i.d. for 26 days (unpaired t test; mean ± SD). Tumor volumes were measured using micro-computed tomography (μCT). Mass of excised tumor tissue is indicated. *p < 0.05; ****p < 0.0001.
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(A) STING transcript levels across TCGA tumor datasets relative to GTEx organ-matched normal tissues. (B) IHC analysis of STING expression in the UCLA PDAC tissue microarray (n = 138). (C) Immunoblot analysis of protein lysates prepared from STING WT and CRISPR-Cas9 KO HS766T or DANG subcutaneous xenograft tumors. (D) IncuCyte live-cell imaging analysis of SUIT2 <t>TetR</t> STINGR284M cells treated +50 ng/mL doxycycline (DOX) ± 500 nM VE-822 (ATR inhibitor [ATRi]) in anchorage-independent cultures (mean ± SD; n = 6; one-way ANOVA corrected for multiple comparisons by Bonferroni adjustment). Images are representative of respective experimental endpoints. (E) Immunoblot analysis of SUIT2 tumors from mice treated ± DOX diet ± AZD-6738 (25 mg/kg b.i.d.) for 3 days. (F) Experimental design to evaluate the impact of ATRi against PDAC tumors with active STING signaling. (G and H) NCG mice were inoculated with subcutaneous (s.c.; G; n = 8) or orthotopic (H; n = 4) SUIT2 TetR STINGR284M tumors. Mice were fed a DOX-supplemented diet starting 7 days after tumor inoculation and treated ±25 mg/kg ATRi AZD-6738 b.i.d. for 26 days (unpaired t test; mean ± SD). Tumor volumes were measured using micro-computed tomography (μCT). Mass of excised tumor tissue is indicated. *p < 0.05; ****p < 0.0001.
Generation Plenti Teton Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


(A) STING transcript levels across TCGA tumor datasets relative to GTEx organ-matched normal tissues. (B) IHC analysis of STING expression in the UCLA PDAC tissue microarray (n = 138). (C) Immunoblot analysis of protein lysates prepared from STING WT and CRISPR-Cas9 KO HS766T or DANG subcutaneous xenograft tumors. (D) IncuCyte live-cell imaging analysis of SUIT2 TetR STINGR284M cells treated +50 ng/mL doxycycline (DOX) ± 500 nM VE-822 (ATR inhibitor [ATRi]) in anchorage-independent cultures (mean ± SD; n = 6; one-way ANOVA corrected for multiple comparisons by Bonferroni adjustment). Images are representative of respective experimental endpoints. (E) Immunoblot analysis of SUIT2 tumors from mice treated ± DOX diet ± AZD-6738 (25 mg/kg b.i.d.) for 3 days. (F) Experimental design to evaluate the impact of ATRi against PDAC tumors with active STING signaling. (G and H) NCG mice were inoculated with subcutaneous (s.c.; G; n = 8) or orthotopic (H; n = 4) SUIT2 TetR STINGR284M tumors. Mice were fed a DOX-supplemented diet starting 7 days after tumor inoculation and treated ±25 mg/kg ATRi AZD-6738 b.i.d. for 26 days (unpaired t test; mean ± SD). Tumor volumes were measured using micro-computed tomography (μCT). Mass of excised tumor tissue is indicated. *p < 0.05; ****p < 0.0001.

Journal: Cell reports

Article Title: Reprogramming of nucleotide metabolism by interferon confers dependence on the replication stress response pathway in pancreatic cancer cells

doi: 10.1016/j.celrep.2021.110236

Figure Lengend Snippet: (A) STING transcript levels across TCGA tumor datasets relative to GTEx organ-matched normal tissues. (B) IHC analysis of STING expression in the UCLA PDAC tissue microarray (n = 138). (C) Immunoblot analysis of protein lysates prepared from STING WT and CRISPR-Cas9 KO HS766T or DANG subcutaneous xenograft tumors. (D) IncuCyte live-cell imaging analysis of SUIT2 TetR STINGR284M cells treated +50 ng/mL doxycycline (DOX) ± 500 nM VE-822 (ATR inhibitor [ATRi]) in anchorage-independent cultures (mean ± SD; n = 6; one-way ANOVA corrected for multiple comparisons by Bonferroni adjustment). Images are representative of respective experimental endpoints. (E) Immunoblot analysis of SUIT2 tumors from mice treated ± DOX diet ± AZD-6738 (25 mg/kg b.i.d.) for 3 days. (F) Experimental design to evaluate the impact of ATRi against PDAC tumors with active STING signaling. (G and H) NCG mice were inoculated with subcutaneous (s.c.; G; n = 8) or orthotopic (H; n = 4) SUIT2 TetR STINGR284M tumors. Mice were fed a DOX-supplemented diet starting 7 days after tumor inoculation and treated ±25 mg/kg ATRi AZD-6738 b.i.d. for 26 days (unpaired t test; mean ± SD). Tumor volumes were measured using micro-computed tomography (μCT). Mass of excised tumor tissue is indicated. *p < 0.05; ****p < 0.0001.

Article Snippet: pLENTI TetR BLAST mammalian expression plasmid , Addgene , Cat#17492.

Techniques: Expressing, Microarray, Western Blot, CRISPR, Live Cell Imaging, Micro-CT

KEY RESOURCES TABLE

Journal: Cell reports

Article Title: Reprogramming of nucleotide metabolism by interferon confers dependence on the replication stress response pathway in pancreatic cancer cells

doi: 10.1016/j.celrep.2021.110236

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: pLENTI TetR BLAST mammalian expression plasmid , Addgene , Cat#17492.

Techniques: Virus, Microarray, Recombinant, Lysis, Protease Inhibitor, Membrane, Saline, Autoradiography, Plasmid Preparation, Control, BIA-KA, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Expressing, Software, Mass Spectrometry, Targeted Proteomics, Real-time Polymerase Chain Reaction, Live Cell Imaging