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. 2020 Oct 16;21(20):7676. doi: 10.3390/ijms21207676

Figure 4.

Figure 4

Canagliflozin ameliorated the upregulation of Spp1 transcriptional activity. (a) Primary culture of proximal tubular epithelial cells (PTEC) derived from enhanced green fluorescent protein (EGFP)-Spp1 knock-in reporter mice. PTEC were cultured in 5 mM (“low”) or 30 mM (“high”) glucose conditions for 7 days. Lactate in the culture supernatants was assessed by enzyme-linked immunosorbent assay (ELISA; n = 6 per group). (b–d) Primary culture of PTEC derived from EGFP-Spp1 knock-in reporter mice. PTEC were cultured in 30 mM glucose conditions for 7 days with 50 mM 2-deoxy-D-glucose (2-DG), 30-µM canagliflozin, 10-µM canagliflozin, or control regent. (b) Lactate in the culture supernatants was assessed by ELISA (n = 6 per group). (c) Analysis of fluorescence microscopy showed 4ʹ,6-diamidino-2-phenylindole (DAPI) and MitoSOX staining and of EGFP-Spp1 expression. (d) Mean fluorescence intensity of MitoSox in PTEC (n = 3–5 per group). (e) Osteopontin (OPN) in the culture supernatants was assessed by ELISA (n = 6 per group). * p < 0.05, ** p < 0.01, *** p < 0.001. Data are shown as means, and error bars depict SEM.