Ferric Ammonium Citrate Upregulates PD-L1 Expression through Generation of Reactive Oxygen Species

Figure 4

Role of ROS in FAC-mediated IL-1β and PD-L1 production. (a) Schematic diagram explaining FAC-mediated inflammatory response in macrophages. (b) BMDMs were incubated with FAC at concentrations of 0.5, 1, and 5 mM for 16 h. ROS production in F4/80⁺ macrophages was measured by flow cytometry. (c, d) BMDMs were incubated with FAC at a concentration of 0.5 mM FAC in the presence of 2 μM ferrostatin, 0.1 mM rotenone, and 0.2 μM NAC for 16 h. ROS (c) and PD-L1 production (d) in F4/80⁺ macrophages were measured by flow cytometry. (e, f) BMDMs were pretreated with LPS 500 ng/mL for 3 h. After priming, 0.5 mM FAC, 2 μM ferrostatin, 0.1 mM rotenone, and 0.2 μM NAC were coincubated with 1 mM ATP for 4 h. ROS (e) and IL-1β production (f) were estimated.

(a)

(b)

(c)

(d)

(e)

(f)