Cut-and-run sequencing of adipose Treg cells with anti-HIF-1α and anti-PPAR-γ antibodies

Molecular mechanisms underlying the development of adipose Tregs remain poorly understood, prompting us to subsequently characterize the critical transcription cascade. The goal of this study is to reveal the function of key transcription factors HIF-1α and PPAR-γ in fate specification of adipose Tregs. Using CUT&RUN assay and sequencing of the enriched DNA, we detected a binding of HIF-1α and PPAR-γ to specific genomic regions in adipose Treg cells. In doing so, we highlighted a potential role of HIF-1α and PPAR-γ in establishing the specific identity of adipose Tregs. Around 30,000 TCRβ+CD4+CD25+YFP+ adipose Treg cells from 4-month old male Foxp3Cre mice were subjected to CUT&RUN assay according to the demonstrated protocol (CUT&RUN, Cell Signaling Technology, Cat#14209). Extracted DNA was subjected to sequencing.