M1 differentiation medium (days 28–29)

Reagent	Final concentration	Amount
Lipopolysaccharides	100 ng/mL	50 μL
IFN-γ	20 ng/mL	10 μL
Basal macrophage M0 medium	N/A	50 mL
Total	N/A	50 mL

Note: Sterilize media using a 0.22 μm membrane filter and store at 4°C for maximum 7 days. Pre-warm the culture medium for 15 min at room temperature before use.