Research Paper Volume 12, Issue 19 pp 19045—19059

FGF23 activates Nrf2 signaling in osteoblasts. OB-6 osteoblastic cells (A–C) or the primary murine osteoblasts (G–I) were treated with FGF23 (25 ng/mL) for 4h, expression of listed genes in total cell lysates was tested by Western blotting and qPCR assays (A, B, G and H), with relative NQO1 activity tested as well (C and I). The control OB-6 cells, pre-treated with LY294002 (500 nM, 30 min pretreatment), as well as the stable OB-6 cells with the lentiviral FGFR1 shRNA (“shFGFR1”) or the CRISPR/Cas9-Akt1-KO construct (“ko-Akt1”), were treated with FGF23 (25 ng/mL) for 4h, expression of HO1 and NQO1 mRNA was shown (D and E). The relative HO1 and NQO1 mRNA expression in stable OB-6 cells with the constitutively-active Akt1 construct (caAkt1) or the empty vector (“Vec”) was tested (F). Data are presented as the mean ± standard deviation (n=5).* p<0.05 vs. PBS treatment. ^# p<0.05. Each experiment was repeated three times and similar results were obtained.