Synthesis and Analysis of TEPP-46 and DASA-58

TEPP-46 and DASA-58 were synthesized in accordance with published methods (12 (link), 13 (link)) and were a kind gift from Craig Thomas, NIH, MD, USA. LPS used was E. coli, serotype EH100 (Alexis). Western blotting antibodies were anti-PKM2 D78A4 (4053) and anti-β-actin (4267) from Cell Signaling Technologies, anti-mB7-H1 (PD-L1, AF1019) from R&D Systems, and anti-HIF-1α (NB100-449) from Novus. Anti-mouse-CD274PE (MIH5) from eBioscience and anti-mouse CD16/CD32 (Mouse BD Fc Block) from BD Pharmingen™ were used for FLOW cytometry in vitro. SYBR primers were from Eurofins, UK. Dimethyl sulfoxide used as vehicle control for TEPP-46 in all relevant in vitro assays. Antibodies used for flow cytometry analysis of in vivo tissues are listed in Table S1 in Supplementary Material.

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Palsson-McDermott E.M., Dyck L., Zasłona Z., Menon D., McGettrick A.F., Mills K.H, & O’Neill L.A. (2017). Pyruvate Kinase M2 Is Required for the Expression of the Immune Checkpoint PD-L1 in Immune Cells and Tumors. Frontiers in Immunology, 8, 1300.

Publication 2017

anti-HIF-1α (NB100-449) anti-mouse CD16/CD32 (Mouse BD Fc Block)

Actin Antibodies Assays Dasa Dimethyl sulfoxide E coli Flow cytometry Mouse Novus Pd l1 Primers Tepp 46 Tissues

Corresponding Organization :

Other organizations : Trinity College Dublin

Top 5 similar protocols

Variable analysis

independent variables

TEPP-46
DASA-58

dependent variables

PKM2 expression
PD-L1 expression
HIF-1α expression

control variables

LPS from E. coli, serotype EH100
Dimethyl sulfoxide (vehicle control for TEPP-46)
Antibodies used for flow cytometry analysis of in vivo tissues (listed in Table S1 in Supplementary Material)

positive controls

Not explicitly mentioned

negative controls

Dimethyl sulfoxide (vehicle control for TEPP-46)

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