Altered biometal trafficking pathways are associated with loss of CLN6 in sheep. (A-F) Densitometry and representative immunoblots of homogenates (5–40 μg) isolated from the occipital lobe of 3, 7 and 14 month old control or CLN6 affected sheep (N = 3 per group) probed with antibodies directed against CLN6 or a range of metal transporters or metal binding proteins. GAPDH, β-tubulin, total Akt or total ERK, as appropriate, were used as loading controls. Quantification was performed in ImageJ and levels are expressed relative to those in control sheep at each age. Data are mean + SEM. *p < 0.05, **p < 0.01, ***p < 0.001 by Student’s t test. C, control. (G-H) Normalized Zip7 protein levels in Merino sheep occipital lobe were plotted against normalized levels of CLN6 (G) or MT (H) to determine correlations between these proteins. Linear regression analysis was performed in GraphPad Prism.