Antihepcidin Spiegelmer inhibits hepcidin effects in vitro. (A) Ferroportin western blot analysis of lysates obtained from mouse J774A.1 macrophages after stimulation with huHep alone or in combination with different concentrations of the L-oligoribonucleotide NOX-H94002 or its PEGylated variant NOX-H94 (1 representative experiment of 9). (B) Ferritin content of lysates obtained from macrophages after stimulation with different concentrations of huHep (1 representative experiment of 3) or (C) after stimulation with 20 nM huHep and different concentrations of NOX-H94. The results from 6 independent experiments were taken to calculate a half maximal inhibitory concentration mean value. Ferritin content was analyzed with a mouse ferritin ELISA (2 lysates, each tested twice in the ELISA, mean ± standard deviation). The amount of ferritin was normalized to total protein.