Fig. 4

Traditional airway surface liquid proteins clustered to fraction 2–4. A Normalized intensities for the molecular extracellular chaperone clusterin (CLU) in unstimulated (open circles, n = 4) and stimulated (filled circles, n = 6) WT piglets (green) and unstimulated (open circles, n = 3) and stimulated (filled circles, n = 3) CF piglets (pink). B There was more clusterin in fraction 4 compared to fraction 1 when normalized intensities from unstimulated and stimulated WT piglets were pooled (n = 10 in each group), ***P = 0.0007, Mann–Whitney test. C Pooled normalized intensities from unstimulated and stimulated WT piglets, fraction 1–4 (n = 10, 40 values) and relative intensities from unstimulated and stimulated CF piglets (n = 6, 24 values) demonstrated that clusterin was higher in CF than WT piglets, *P = 0.02, Mann–Whitney test. D The serine protease inhibitor alpha-1 antitrypsin (SERPINA1) was found in WT piglets (green), fraction 1–4 but hardly at all in fraction 5, unstimulated samples (open circles, n = 4) and stimulated samples (filled circles, n = 6) and displayed the same pattern in CF piglets (pink) as in WT piglets, unstimulated samples (open circles, n = 3) and stimulated samples (filled circles, n = 3). E There was more alpha-1 antitrypsin in fraction 1–4 from CF piglets than fraction 1–4 from WT piglets, pooled values from unstimulated and stimulated piglets (WT n = 10, 40 values, CF n = 6, 24 values), **P = 0.009, Mann–Whitney test. F Orosomucoid 1 (ORM1) displayed a trend for increasing levels from 1–4 in WT tracheas (green), unstimulated (open circles, n = 4) and stimulated (filled circles, n = 6) and a similar trend in CF piglets (pink), unstimulated (open circles, n = 3) and stimulated (filled circles, n = 3). G There was more orosomucoid 1 in fraction 1–4 from CF piglets than fraction 1–4 from WT piglets, pooled relative intensities from unstimulated and stimulated piglets (WT n = 10, 40 values, CF n = 6, 24 values), *P = 0.02, Mann–Whitney test. Each data point in A, B, D and F represents one piglet and results are presented as median with interquartile range. H Immunostaining of paraffin sections from newborn WT piglets visualized orosomucoid 1 (red) collected on the airway surface (dashed line). Scale bar: 5 µm. I Immunostaining of paraffin sections from newborn WT piglets. Orosomucoid 1 (red) was detected in small capillaries (CD31, green) in the airway submucosa, indicated by white arrow and dashed line. Scale bar in H and I 10 µm