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. 2012 Jul 3;303(5):F721–F732. doi: 10.1152/ajprenal.00157.2012

Fig. 7.

Fig. 7.

PD-81723 induces sphingosine kinase (SK)1 expression and activity in HK-2 cells. A and B: representative images (top) and relative normalized band intensities (bottom) for SK mRNA (by RT-PCR; A) and protein (by immunoblot analysis; B) expression in HK-2 cells treated with vehicle (1% DMSO) or 10 μM PD-81723 (n = 4 per group). HK-2 cells were treated for 6 h (mRNA analysis) or 16 h (protein analysis). Note that PD-81723 increased SK1 mRNA and protein expression in HK-2 cells but did not change SK2 expression. C: SK activity in HK-2 cells treated with vehicle or with 1–10 μM PD-81723 6 h (n = 4 per group). PD-81723 caused a dose-dependent increase in SK activity in HK-2 cells. We preferentially measured SK1 activity by the addition of Triton X-100 in our SK activity assay. A selective A1AR antagonist (100 nM DPPCX) significantly attenuated the PD-81723-induced increase in SK activity. Values are means ± SE. *P < 0.05 vs. vehicle; #P < 0.05 vs. PD-81723.