Effect of androgen receptor signaling on H3K27 trimethylation in Th17 cells
Females have increased prevalence of many Th17-mediated diseases, including asthma. Androgen signaling decreases Th17-mediated airway inflammation, and Th17 cells rely on glutaminolysis. However, it remains unclear whether androgen receptor (AR) signaling modifies glutamine metabolism to suppress Th17-mediated inflammation. In this experiment, we conducted a CUT & RUN to determine how AR signaling modified histone 3 lysine 27 trimethylation in Th17 cells from WT males, WT females, and ArTfm male mice (n=3 from each group). Overall design: Naïve T cells were isolated from spleens of WT male, WT female, and ArTfm male mice. ArTfm mice have a mutation in the androgen receptor (AR) that makes it non-functional.  Th17 cells were differentiated in culture for 4 days. Cells were isolated and using a H3K27me3 antibody or IgG isotype control and DNA was extracted using the KAPA Hyper Prep kit protocol. Sequencing libraries were constructed adn sequenced for 150 cycles in paired-end mode on Ilumina Nova-Seq 6000 platform.