DataSheet_1_Ehf and Fezf2 regulate late medullary thymic epithelial cell and thymic tuft cell development.pdf

Thymic epithelial cells are indispensable for T cell maturation and selection and the induction of central immune tolerance. The self-peptide repertoire expressed by medullary thymic epithelial cells is in part regulated by the transcriptional regulator Aire (Autoimmune regulator) and the transcription factor Fezf2. Due to the high complexity of mTEC maturation stages (i.e., post-Aire, Krt10+ mTECs, and Dclk1+ Tuft mTECs) and the heterogeneity in their gene expression profiles (i.e., mosaic expression patterns), it has been challenging to identify the additional factors complementing the transcriptional regulation. We aimed to identify the transcriptional regulators involved in the regulation of mTEC development and self-peptide expression in an unbiased and genome-wide manner. We used ATAC footprinting analysis as an indirect approach to identify transcription factors involved in the gene expression regulation in mTECs, which we validated by ChIP sequencing. This study identifies Fezf2 as a regulator of the recently described thymic Tuft cells (i.e., Tuft mTECs). Furthermore, we identify that transcriptional regulators of the ELF, ESE, ERF, and PEA3 subfamily of the ETS transcription factor family and members of the Krüppel-like family of transcription factors play a role in the transcriptional regulation of genes involved in late mTEC development and promiscuous gene expression.</p

Supplementary Figure S1 from Targeting Cytokine Therapy to the Pancreatic Tumor Microenvironment Using PD-L1–Specific VHHs

S1. B3 binds to PD-L1 on multiple tumor types.</p

Supplementary Figure Legends from Targeting Cytokine Therapy to the Pancreatic Tumor Microenvironment Using PD-L1–Specific VHHs

Supplementary Figure Legends from Targeting Cytokine Therapy to the Pancreatic Tumor Microenvironment Using PD-L1–Specific VHH

Supplementary Figure S3 from Targeting Cytokine Therapy to the Pancreatic Tumor Microenvironment Using PD-L1–Specific VHHs

S3. Single agent treatment with B3 does not alter B16 growth in vivo.</p

Supplementary Figure S5 from Targeting Cytokine Therapy to the Pancreatic Tumor Microenvironment Using PD-L1–Specific VHHs

S5. Combination B3-IL2 and A12-IFNγ has additive benefit over either agent alone.</p

Supplementary Figure S4 from Targeting Cytokine Therapy to the Pancreatic Tumor Microenvironment Using PD-L1–Specific VHHs

S4. Lack of efficacy of B3-IL2 in KPC model and A12-GMCSF in Panc02 model.</p

Supplementary Figure S2 from Targeting Cytokine Therapy to the Pancreatic Tumor Microenvironment Using PD-L1–Specific VHHs

S2. In vitro killing of IFNγ treated B16-ova cells by activated OT-I T cells is inhibited by PD-L1.</p

Supplementary Figure S6 from Targeting Cytokine Therapy to the Pancreatic Tumor Microenvironment Using PD-L1–Specific VHHs

S6. A12-IFNγ alters the tumor microenvironment in KPC organoid model of pancreatic cancer.</p