Fig. 1: Renal copper and CTR1 are markedly increased in renal fibrosis.

a Kidney copper, zinc, magnesium, manganese, iron, chrome and calcium concentrations were determined by ICP-MS analysis on day 28 after uIRIx operation (n = 6). b ICP-MS analysis of copper concentrations in sham and UUO kidneys on day 7 (n = 8). c ICP-MS analysis showing the copper concentrations in NRK-49F cells after 48 h of TGF-β1treatment (n = 5). d Immunofluorescent co-staining on sham and UUO kidney sections showed that CTR1 and α-SMA were increased in UUO kidney sections (n = 3). Bar = 50 μm. e Rat kidney sections were stained with CTR1 and Pan Cytokeratin antibody (n = 3). Bar = 50 μm. f Representative western blotting analysis showing the expression of CTR1 in sham and UUO kidneys on day 7 (n = 8). g The CTR1 shRNA–expressing plasmid or the empty plasmid mixed with lipid microbubble were delivered into the rat kidneys immediately after the UUO operation through the ultrasound-mediated gene transfer technique. Renal copper concentrations were tested by ICP-MS (n = 8). h Western blotting showing CTR1 upregulation in NRK-49F cells treated with TGF-β1 (n = 3). i CTR1 and α-SMA were analyzed by immunofluorescent co-staining in NRK-49F cells with or without TGF-β1 treatment (n = 3). Bar = 50 μm. j ICP-MS was used to analyze the copper concentrations in NRK-49F cells after CTR1 downregulation by lentivirus (n = 5). Data represent the mean ± SEM. **P < 0.01, ***P < 0.001 versus sham-operated mice. ##P < 0.01, ###P < 0.001 versus sham-operated rats. $$P < 0.01 versus the nontreated group. &P < 0.05 versus with the scramble-treated UUO kidneys. ¥P < 0.05 versus TGF-β1-treated cells.