Research Perspective Volume 1, Issue 8 pp 740—745

Figure 3. IL-8 protein concentration in culture medium measured by ELISA. ARPE-19 cells were treated with 150 uM H₂O₂ in culture medium with 10% fetal bovine serum for 2 hours and allowed to recover in stressor-free ARPE medium for 22 hours. The procedure was repeated to generate the next treatment cycle. The twice treated cells were allowed to stay in 1% serum ARPE medium for 72 hours after stress before proceeding to further analytic assays. The culture media from control and senescent RPE cells were collected and used directly for ELISA measurement. IL-8 secretion level was measured in pg/ml using human IL-8 ELISA Kit (BioLegend, Inc., San Diego, CA) according to manufacturer's instructions. The level of IL-8 secretion shown here was averaged from a triplicate of each sample and from 3 independent repeats of H₂O₂ treatments. Student's t test was used for statistical analysis (**; p < 0.0005).