Fig. 6.

Validation analysis in human glomerular endothelial cells (GECs) and rat kidney tissues for the identified chronic injury markers, protein S and galectin-1.A, western blot analysis was performed for fibronectin, phospho-Ser536-P65, protein S, and galectin-1 at 24 and 48 h after hypoxic injury in GECs. Representative western blot (upper) and quantification of the blots (lower). Asterisks in the densitometry are calculated by comparison with the levels in normoxic GECs. Error bars represent standard deviation. ∗p < 0.05; ∗∗p < 0.005; ∗∗∗p < 0.001. B, mRNA expression of PROS-1 and LGALS-1 was measured after 24 and 48 h of hypoxic injury in GECs and compared with the mRNA expression in normoxic GECs. Error bars represent standard deviation. ∗p < 0.05; ∗∗∗p < 0.001. C, periodic acid–Schiff stain, Masson's trichrome stain, and immunohistochemical stain for protein S and galectin-1 of the 5/6 nephrectomized rat kidney tissue at 4 and 8 weeks after injury. Original magnification, ×200. Scale bars: 100 μm. D, western blot analysis was performed for vimentin, phospho-Ser536-P65, protein S, and galectin-1 in rat kidney tissues at 4 and 8 weeks after 5/6 nephrectomy. Asterisks in the densitometry are calculated by comparison with the levels of sham-operated rat kidney tissue. Error bars represent standard deviation. ∗p < 0.05; ∗∗p < 0.005; ∗∗∗p < 0.001. E, PROS-1, LGALS-1, NFKB1, P53, and fibronectin mRNA levels were measured in sham-operated rat kidney and 5/6 nephrectomized rat kidney at 4 and 8 weeks. Error bars represent standard deviation. ∗p < 0.05; ∗∗p < 0.005; ∗∗∗p < 0.001.