Fig. 4.

Effect of tandem addition of rotenone and LPS on dopaminergic neurodegeneration. A, Tandem addition without change of medium in between. Rat neuron–glia cultures were treated for the indicated periods with rotenone (0.5 nm) and/or LPS (0.5 ng/ml) in the following manner: rotenone or LPS for the entire 8 d (0–8) or only the last 4 d (4–8), rotenone for 8 d (0–8) plus LPS for the last 4 d (4–8), or LPS for 8 d (0–8) plus rotenone for the last 4 d (4–8). Afterward, neurotoxicity was determined by DA uptake and quantification of TH-IR neurons. **p < 0.005 compared with the cultures treated with rotenone alone;⁺p < 0.05 and⁺⁺p < 0.005 compared with the cultures treated with LPS alone. B, Tandem addition with change of medium. Rat neuron–glia cultures were treated for the indicated periods of time with vehicle, 0.5 nm rotenone, and/or 0.5 ng/ml LPS in sequential orders and with medium changes in between as follows: rotenone or LPS alone for the first 3 d only (0–3) or the last 3 d only (5–7), rotenone for the first 3 d (0–3) followed by resting for 1 d and then LPS for 3 d (5–7), LPS for the first 3 d (0–3) followed by resting for 1 d and then rotenone for 3 d (5–7). Afterward, neurotoxicity was determined by DA uptake and quantification of TH-IR neurons. The results are the mean ± SEM of two experiments performed in triplicate. *p < 0.05 compared with the cultures treated with rotenone alone;⁺p < 0.05 compared with the cultures treated with LPS alone. Rot, Rotenone.