Figure 1
From: Bone marrow stromal cell-derived hepcidin has antimicrobial and immunomodulatory activities
Hepcidin is expressed in BMSCs and is inducible upon microbial and inflammatory stimuli. (A) PCR showing human hepcidin gene (HAMP) expression in BM stromal cells (BMSCs). HAMP primer product size is 201 bp. (B) PCR showing mouse hepcidin gene (Hamp) expression. Hamp primer product size is 188 bp. (C) Dot blot of wild type and Hamp−/− mice livers to show specificity of the antibody used. (D) Immunofluorescent detection of hepcidin in BMSC isolated from Hamp−/− and WT mice or obtained from human volunteers (nuclear staining in blue and hepcidin in red). Scale bars represent 10 µm (mouse) and 16 μm (human). (E) Hepcidin mRNA expression in unstimulated hBMSCs, IL-6- and BMP2-stimulated hBMSCs, human liver and THP-1 macrophage cells [compared to housekeeping (hkg) control gene expression] (F–H): Human BMSC HAMP expression in response to 6 h (F) TNF-α, (G) INF-γ and (H) IL-6 stimulation at different doses [“c” (control, no cytokine), 1 ng/mL and 10 ng/mL]. (I) HAMP mRNA expression in control hBMSCs or hBMSCs stimulated with Pseudomonas aeruginosa for 4 or 9 h. Human hepcidin Taqman qPCR results were normalized to beta-actin expression. (J, K) Human BMSC cultures from five independent donors were left unstimulated or were stimulated with 200 ng/mL BMP2 or 50 ng/mL IL-6 for 24 h in vitro. Hepcidin concentrations were determined by ELLA in (J) collected and concentrated SNs and (K) cell lysates. One-way ANOVA, *p < 0.05; **p < 0.01; ***p < 0.001.
