FIGURE 5.

IL-1R1 on CD11c⁺ BMDC generates IL-1ra to limit IL1β-induced renal tubular cell injury in vitro. Myel KO GM-CSF-derived bone marrow dendritic cells (BMDCs) produce less anti-inflammatory Il1rn in response to IL1β which induce increased HK2 kidney tubular cellular injury, and IL-1ra (Anakinra) treatment reverses the heightened HK2 cellular injury. (A) Schematic experimental workflow for induction of BMDCs from Myel WT and Myel KO mice (Created with Biorender.com). (B) Il1r1 mRNA expression in Myel WT and Myel KO BMDC. (C) Il1rn mRNA expression in Myel WT and Myel KO BMDCs upon IL1β (Created with Biorender.com) or (D) LPS stimulation (Created with Biorender.com) (n = 4; ***p < 0.001). (E) HK2 kidney tubular cells were co-cultured with BMDCs from Myel WT or Myel KO mice in transwell system for 12 h (Created with Biorender.com). (F) NGAL (LCN2) mRNA expression was measured in HK2 cells. Dots represent individual samples from each group. Statistical analysis performed by two-way ANOVA with Sidak test. (n = 3; **p < 0.01). (G) Diagram of experimental workflow. HK2 cells were treated with cell medium harvested from either vehicle or IL1β-stimulated Myel WT or Myel KO BMDCs. Media was subsequently treated with vehicle or recombinant IL-1Ra (Anakinra) (Created with Biorender.com. (H) Shown in graph is fold change in Ngal mRNA from HK2 cells. Significance was determined by two-way ANOVA with Sidak test (n = 6; ****p < 0.0001).