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Figure 1

From: Role of the kidneys in the redistribution of heme-derived iron during neonatal hemolysis in mice

Figure 1

Markers of hemolysis in the plasma of mouse neonates. (a) Total bilirubin concentration in the plasma of mouse neonates and adult (3-month-old) mice. Values are expressed as the means ± S.D. N values for each group are: 3dpp = 5, 5dpp = 5, 7dpp = 5, 9dpp = 5, 11dpp = 5, 90dpp = 5. Data set is not normally distributed, therefore, nonparametric, Kruskal-Wallis ANOVA was used (p = 0,004, df = 5, H = 17,3). Dunn’s Multiple Comparison Test was used as post-hoc test. Plasma levels of hemopexin (Hpx) (b) and haptoglobin (Hp) (d) assessed by Western blotting. The blots were reprobed with polyclonal anti-albumin antibody as a loading control. Results come from 2 separate blots and relative levels of proteins were standardized to a percentage with %S.D. in order to eliminate technical differences between blots. Immunolabeled Hpx (c) and Hp (e) and albumin control bands were quantified using a Molecular Imager and the relative levels of the test proteins (means ± S.D.) are plotted in arbitrary units. N values in each group for both Hp and Hpx are: 3dpp = 5, 5dpp = 5, 7dpp = 5, 9dpp = 5, 11dpp = 4. Data set for Hpx has normal distribution, therefore, one-way ANOVA was used (p < 0.0001, df = 4, F = 51,40). Tukey’s Multiple Comparison Test was used as post-hoc test. Data set for Hp is not normally distributed, therefore, Kruskal-Wallis ANOVA was used (p = 0,9622, df = 4, H = 0,608) but no statistically significant differences were observed. Capital and small letters over bars in the charts (a,c,e) denote significant differences between age groups, with p < 0.01 and p < 0.05, respectively. dpp – days postpartum. (fh) RBC indices potentially indicating the occurrence of hemolysis: (f) RBC- Red blood cells count, (g) MCV - mean corpuscular volume of a red blood cell, (h) RDW - red cell distribution width. Values are expressed as the means ± S.D. N values for each group are: 3dpp = 6, 5dpp = 5, 7dpp = 5, 9dpp = 4, 11dpp = 7. Data set for RBC and RDW has normal distribution, therefore, one-way ANOVA was used and Tukey’s Multiple Comparison Test was used as post-hoc test for the RBC and post test for linear trend (p = 0,0097) was used for RDW. Data set for MCV is not normally distributed, therefore, nonparametric, Kruskal-Wallis ANOVA was used. Dunn’s Multiple Comparison Test was used as post-hoc test. Capital and small letters over bars in the charts (a,c,f,g,h) denote significant differences between age groups, with p < 0.01 and p < 0.05, respectively. dpp – days postpartum. (i) Lactate dehydrogenase (LDH2) plasma levels in the serum from all experimental group of neonates analyzed by Western blotting as described in Materials and Methods. Lack of LDH2-positive band in the serum of the adult, intact mouse. Prominent immunopositive band is visible in the serum from adult phenylhydrazine-treated (PHZ) mouse (positive control). The blots were reprobed with polyclonal anti-albumin antibody as a loading control.

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