For ATP assays, BMDMs were plated at 5×105/mL and were lysed with Luciferase Cell Culture Lysis Reagent (Promega, Madison, WI). ATP concentration in the lysate was the incubated with firefly luciferase along with the luminescent ATP Determination Kit (Invitrogen, Carlsbad, CA). Luminescence was then determined by the BioTek Synergy MX plate reader (Biotek, Winooski, VT).
For glucose and lactate measurements, glucose and lactate in the media were measured by the YSI 2300 Stat Plus Glucose & Lactate Analyzer (YSI, Yellow Springs, OH). For nitric oxide secretion assays, nitrites in the media were detected by Griess Reagent Kit (Thermo Fisher Scientific, Waltham, MA). Media was incubated with a 1:1 mixture of N-(1-naphthyl)ethylenediamine and sulfanilic acid for 30 minutes at room temperature and were assessed by spectrophotometric analysis on the BioTek Synergy MX plate reader (Biotek, Winooski, VT). For Mito Tracker staining, MitoTracker Green was purchased from Thermo Fisher (Waltham, MA). BMDMs were incubated in MitoTracker Green for 45 minutes and then assessed via flow cytometry.
For glucose and lactate measurements, glucose and lactate in the media were measured by the YSI 2300 Stat Plus Glucose & Lactate Analyzer (YSI, Yellow Springs, OH). For nitric oxide secretion assays, nitrites in the media were detected by Griess Reagent Kit (Thermo Fisher Scientific, Waltham, MA). Media was incubated with a 1:1 mixture of N-(1-naphthyl)ethylenediamine and sulfanilic acid for 30 minutes at room temperature and were assessed by spectrophotometric analysis on the BioTek Synergy MX plate reader (Biotek, Winooski, VT). For Mito Tracker staining, MitoTracker Green was purchased from Thermo Fisher (Waltham, MA). BMDMs were incubated in MitoTracker Green for 45 minutes and then assessed via flow cytometry.