To address what marker(s) is/are most suitable for determining renal cell senescence, cell area, granularity, cycle shift/arrest, SA-β-Gal, SIRT1 and p16 were evaluated after inducing senescence in HK-2 cells with 0.2-0.8 mM HO. Only cell area and granularity concentration-dependently increased at all time-points, whereas SA-β-Gal, SIRT1 and p16 showed significant coefficient of determination (R) at two time-points. Cell granularity had significant correlation coefficient (R) with other six, whereas SA-β-Gal had significant R with five, and cell area, SIRT1 and p16 had significant R with four others. Comparing to SA-β-Gal, other markers had significantly lower fold-changes only at 72-h with 0.8 mM HO, whereas p16 provided greater fold-changes at 48-h with 0.4 and 0.8 mM HO. Therefore, cell area, granularity, SA-β-Gal and p16 may serve as the most suitable markers for determining HO-induced senescence in HK-2 renal cells, whereas other markers can be also used but with inferior quantitative precision.
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