Figure 2.

Establishment of the maximum-tolerated dose of NAC for cultured cells. A: RPE19α, RCF, and FαΔx cells were plated in complete growth medium at 3 × 10⁴ cells/well of a 24-well plate. Within 24 hours, the indicated concentration of NAC was added. The media were refreshed daily, and the experiment was continued for a total of seven days. The pictures shown were taken on day seven. In the presence of 50 mmol/L NAC, nuclear condensation developed in all three cell types. Scale bar = 100 μm. B: The cells described in A were counted on day seven, and the data (cell numbers) from three independent experiments were graphed (the mean ± SD). Blue, red, yellow, and green correspond to 0, 10, 20, and 50 mmol/L NAC, respectively. C: The cells described in B were replated in complete growth medium, and the number of attached cells was counted at the 6-hour time point. The data are expressed as a percentage of re-attached cells and are shown as an average ± SD from three independent experiments. Black, gray, blue, and green correspond to 0, 10, 20, and 50 mmol/L NAC, respectively, which was the concentration of NAC in the culture medium before replating.