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All tumor measurements and calculations were performed by trained technicians blinded to experimental conditions. All images were acquired using a 30-s pfizer my world time and low binning.

Analysis was performed using Living Image software (PerkinElmer). PET analysis was performed with either Pfizer my world software (version 3. A volume of interest was drawn on whole tumors, and the maximum standard uptake value (SUVmax) was recorded. Radioactive probe pfizer my world assays were conducted as previously described (62). Following incubation, cells were washed three times with 1 mL PBS and lysed using radioimmunoprecipitation assay (RIPA) buffer.

Cell lysate radioactivity was measured on a Wizard2 gamma counter (PerkinElmer). Blood was collected in lithium heparin-coated tubes using the retro-orbital technique and stored on ice until plasma isolation. Fragments from resected tumors were weighed (30 to 80 mg), transferred to Omni Hard Tissue homogenization vials, and snap pfizer my world. Immunoblot analysis was performed as previously described (29).

HRP was activated by incubating membranes with a mixture of SuperSignal Pico and SuperSignal Femto enhanced chemiluminescence reagents (100:1 ratio). HRP signals were detected by exposure of autoradiography film or imaging using a LI-COR Odyssey system.

Antibodies are reported in Key Pfizer my world Table (SI Appendix). Total RNA was isolated from cell cultures using the NucleoSpin RNA kit. Reverse transcription was performed using the High Capacity cDNA Reverse Transcription kit. Primer sequences are reported in Key Resource Table. Images were acquired at 3 h intervals over the indicated time period.

Sphere area analysis was applied to quantify johnson 18v. For generation of stable knockdown cell lines, PDAC cells were steps of research with lentivirus harvested from FT293 cells in the presence of polybrene.

Lentivirus-containing supernatants were passed through a 0. Following transduction, cells underwent antibiotic selection, and pfizer my world efficiency was confirmed using immunoblot analysis. All guide RNA (gRNA)-encoding oligonucleotides were cloned into the LentiCrispr version 2 vector. Lipofectamine 3000 was used to transfect Pfizer my world cells with gRNA-specific LentiCrispr version 2 vectors.

Following puromycin selection, cells were singly cloned, and gene knockout was confirmed by genomic DNA PCR and tracking of insertions and deletions by decomposition (TIDE) analysis of Sanger sequencing results. Gene knockout was additionally validated using immunoblot analysis. The generation of SUIT2-TetR-STINGR284M cells was previously described (19). For virus production, lentivirval vectors and packaging plasmids (psPAX2 and pMD2G) at a 2:1:1 ratio were transfected into FT293 cells using polyethylenimine.

Transduced cells were selected in puromycin for 1 wk. Genomic DNA was extracted as previously described using the Pfizer my world Quick-genomic DNA MiniPrep kit and hydrolyzed to nucleosides using the DNA Degradase Plus kit, following manufacturer-supplied instructions (29). Analysis of hydrolyzed DNA, media, serum, and tumor nucleoside levels was performed as previously described (27).

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