<i>L. amazonensis</i> infection inhibits Fpn1 expression in macrophages in a TLR4 dependent manner.

<p>BMDM were left untreated (Control) or incubated with Fe-NTA overnight, washed and infected or not with <i>L. amazonensis</i> axenic amastigotes (<i>L.a</i>) for 1 h. BMDM were washed to remove extracellular parasites, cultured at 34°C for another 24 or 48 h and <i>Fpn1</i> or hypoxantine phosphoribosyltransferase (<i>HPRT1</i>) mRNA was quantified by qPCR. (<b>A</b>) Effect of <i>L. amazonensis</i> infection on ferroportin (<i>Fpn1</i>) mRNA in wild type (WT) BMDM. (<b>B</b>) Effect of <i>L. amazonensis</i> infection on ferroportin (<i>Fpn1</i>) mRNA in TLR4^−/− BMDM. The values represent <i>Fpn1</i> mRNA levels normalized to <i>HRPT1</i> (mean +/− SD of triplicates). (**) p<0.01; (***) p<0.001 (unpaired Student's t test). (<b>C</b>) Fpn1 protein levels in WT and TLR4⁻/⁻ BMDM treated as in (A, B) and infected or not with <i>L. amazonensis</i> axenic amastigotes (<i>L.a</i>) for 48 h. BMDM lysates were analysed by Western Blot with anti-Fpn1 antibodies. Anti-actin antibodies were used as loading controls. Fpn1/actin ratios were determined by quantitative digital imaging. The data are representative of at least 2 independent experiments.