Fig. 2.

Soluble α-Klotho (s-KL) was downregulated in osteogenic-induced human renal interstitial fibroblasts (hRIFs) and HK-2 cells. A Alizarin Red staining (ARS) for calcium deposits in hRIF layers induced with normal medium (NM) for 14 days; n = 3. B Alizarin Red staining (ARS) for calcium deposits in hRIF layers cultured with osteogenic medium (OM) for 14 days; n = 3. C Representative western blots of s-KL and osteogenic markers (OCN, MSX2, Runx2) in hRIFs 1, 4, 7, and 14 days after culture in either OM or NM; n = 3. D Relative alkaline phosphatase (ALP) activity in hRIF lysates 1, 4, 7, and 14 days after culture in either OM or NM; n = 3. E–G Correlation analysis between the relative protein expression levels of s-KL and osteogenic markers in OM-induced hRIFs. H ARS for calcium deposits in HK-2 cell layers cultured with NM for 14 days; n = 3. (I) ARS for calcium deposits in HK-2 cell layers induced with OM for 14 days; n = 3. J Representative western blots of s-KL and osteogenic markers in HK-2 cells 1, 4, 7, and 14 days after culture in either OM or NM; n = 3. K ALP activity in HK-2 cell lysates 1, 4, 7, and 14 days after culture in either OM or NM; n = 3