description: Protocols: C57BL/6J mice were used in this experiment. Dissociation according to Miltenyi mouse lung dissociation kit (https://www.miltenyibiotec.com/DE-en/products/lung-dissociation-kit-mouse.html#gref). The Miltenyi proprietary protocol of the kit was strictly followed. Debris were removed using the debris removal solution by Miltenyi, following strictly the Miltenyi protocol (https://www.miltenyibiotec.com/DE-en/products/debris-removal-solution.html#gref). No particular cell types were enriched or filtered out. There were 2-3 mice per experimental group, which were pooled for each sample. For all studies, mice were randomized into treatment groups according to tumor volume on the day of treatment initiation (day 0). Bintrafusp alfa, anti-PD-L1, TGF-β trap, and isotype control Bintrafusp alfa (BA, previously reported as M7824) is a bifunctional fusion protein composed of the extracellular domain of the TGF-βRII to function as a TGF-β “trap” fused to a human IgG1 antibody blocking PD-L1. The isotype control is a mutated version of anti-PD-L1, which completely lacks PD-L1 binding. In tumor-bearing mice, BA (492 μg) or isotype control (400 μg) were administered with an intravenous injection (i.v.) in 0.2 mL PBS. Mice were treated with 15 Gy RT and, 16 weeks later, treated with Bintrafusp (492 μg) or isotype control (400 μg) on day 0, 3, and 6. Radiation (RT) To assess the combination of radiation with Bintrafusp (BA), mice were randomized into the following treatment groups: isotype control (400 μg), RT (exact dose and schedule provided in the figure legends) + isotype control, BA (492 μg), or BA + RT (BART). Mice were treated with 15 Gy RT 16 weeks prior to BA or isotype control. Whole thoracic irradiation was administered. Briefly, a photon beam was delivered by a 6 MV Artiste Linac (Siemens) at a dose rate of 3 Gy/min in anesthetized mice. The LINAC generates a (6 MV) x-ray photon beam with a dose rate of 3 Gy/min. According to 10x genomics proprietary protocol of the kit According to 10x genomics proprietary protocol of the kit: https://support.10xgenomics.com/single-cell-gene-expression/library-prep/doc/user-guide-chromium-single-cell-3-reagent-kits-user-guide-v2-chemistry) page31ff
attributes.tag: ENA first public
attributes.value: 2021-08-10
attributes.tag: ENA last update
attributes.value: 2021-08-10
attributes.tag: External Id
attributes.value: SAMEA9327043
attributes.tag: INSDC center alias
attributes.value: Translational Radio Oncology (E210)
attributes.tag: INSDC center name
attributes.value: Translational Radio Oncology (E210)
attributes.tag: INSDC first public
attributes.value: 2021-08-10T00:22:25Z
attributes.tag: INSDC last update
attributes.value: 2021-08-10T00:22:25Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10633:BART2_172L_new
attributes.tag: age
attributes.value: 31
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: organism part
attributes.value: lung
attributes.tag: sample name
attributes.value: E-MTAB-10633:BART2_172L_new
attributes.tag: strain
attributes.value: C57BL/6J