Fig 1.
Inflammatory status of the CFTR KO mice.
WBC (white blood cells), lymphocytes, monocytes and neutrophils were measured in the blood of WT and CFTR KO mice (A). Cytokines levels were measured by the V-PLEX Proinflammatory Panel1 (mouse) Kit in the plasma of WT and CFTR KO mice according to the manufacturer’s instruction (B). Duodenal mRNA levels relative to cyclophilin-A expression were assessed by real-time PCR for the indicated genes (C). Data are presented as mean ± SEM. * P<0.05.
Fig 2.
Red cells, iron indices, and iron-related genes in CFTR KO mice.
Hematological and iron parameters were analyzed in WT and CFTR KO mice: RBC (red blood cells), Hb (hemoglobin), HC (hematocrit) and MCV (mean corpuscular volume) (A); plasma iron, TfS, (transferrin saturation) and ferritin (B). Liver hepcidin1 and BMP6 mRNA levels relative to cyclophilin-A were analyzed by real-time PCR (C). Data are presented as mean ± SEM. * P<0.05.
Fig 3.
Lung iron phenotype in CFTR KO mice.
Representative images of BAL (bronchoalveolar lavages) cytospin slides obtained from WT and CFTR KO mice and stained with Perl's Prussian blue. Iron loaded macrophages are indicated by arrows. Magnification X40 (A). Perls’ blue staining of lung section from WT and KO mice (B). Lung iron content (C) and L-ferritin analysis from cytosolic fractions; the right panel represents the quantification of the blots (arbitrary units) (D). Data are presented as mean ± SEM.
Fig 4.
Duodenal and spleen iron-related proteins in CFTR KO mice.
Duodenum (A) and spleen (B, C) were collected from WT and CFTR KO mice. Duodenum DcytB, DMT1 and ferroportin were analyzed using proteins from membrane enriched-fractions and L-ferritin from cytosolic fractions. The loading control, ß-actin, is shown (A). Perls’ blue staining of spleen section from WT and KO mice (B). Spleen ferroportin was analyzed using proteins from membrane enriched-fractions and L-ferritin from cytosolic fractions. The loading control, ß-actin, is shown. The right panel represents the quantification of the blots (arbitrary units). Data are presented as mean ± SEM. * P<0.05.
Fig 5.
Erythrophagocytosis in presence of CFTR(inh)-172.
Two independent experiments (Exp1, 2) of erythrophagocytosis (EP) using normal or aged red blood cells were performed with bone-marrow derived macrophages incubated or not with 10-5M CFTR(inh)-172.