Fig. 5.

Effects of hypoxia and hypoxia mimetics on FcγRIIb expression and its transcriptional regulation. a, Histograms showing expression of FcγRIIb on LD and HD monocytes cultured under 21% or 1% O₂, b, and quantified using flow cytometry (n = 5 per group). c, Histograms showing expression of FcγRIIb on monocytes treated with DMOG or Roxadustat (Rox). d, and quantified using flow cytometry (n = 5–10 per group). e, FcγRIIb expression quantified using flow cytometry following dose titration of DMOG treatment of THP-1 cells (n = 3, bars show means ± SEM). f, FcγRIIb expression on untreated (U) and DMOG treated M0, M1 and M2 monocyte-derived macrophages (MDM), (n = 11 per group). g, Representative Western blot showing FcγRIIb expression in untreated or DMOG-treated (D) M1 macrophages for 2 donors. h, FcγR A:I ratio on monocytes, THP-1 cells and MDMs untreated or treated with DMOG or Roxadustat (Rox) (n = 5–11 per group). i, FcγRIIb expression (left) and FcγR A:I ratio (right) on monocytes following treatment with VH298 (n = 8). j, FcγRIIb expression (left) and FcγR A:I ratio (right) on DMOG-treated monocytes in the absence or presence of FM19G11 (FM19, n = 8). k, Representative Western blot showing c-Jun expression in Untreated (U) or DMOG-treated (D) monocytes l, combined Western blot data of fold change of c-Jun expression relative to HSC70 (n = 6). m, FcγRIIb expression and n, FcγR A:I ratio on DMOG-treated monocytes following c-Jun peptide treatment (c-Jun; n = 8). o, Representative histograms showing FcγRIIb expression on purified human untreated (U) monocytes transfected with scrambled control (Scram) siRNA, and DMOG-treated monocytes transfected with Scram, HIF2A or JUN siRNA, 24 h post-treatment. p, FcγRIIb expression and q, FcγR A:I ratio for 9–17 donors per group using flow cytometry following treatments stated in o. r, Representative Western blots showing FcγRIIb expression on untreated monocytes transfected with Scram siRNA, and DMOG-treated monocytes transfected with Scram, HIF2A or JUN siRNA. s, Combined Western blot data of fold change of FcγRIIb expression relative to HSC70 (n = 9). Each point on the graphs represents a unique donor and bars represent group means. Statistical significance was assessed using a paired two-tailed Wilcoxon test (*p < 0.05, **p < 0.01, ****p < 0.0001 and ns = non-significant). Also see Additional file 5: Fig. S5