Figure 1.

Induction of Alox5 by MWCNTs. (A and B) J774A.1 macrophage cells were untreated or treated with control media (DMEM + 1% FBS), MWCNTs (2.5 or 10 μg/ml), carbon black (2.5, 10, or 30 μg/ml), or M1 inducer (IFN-γ at 20 ng/ml plus LPS at 100 ng/ml) for 1 day (A) or 3 days (B). Total RNA was isolated and mRNA levels of Alox5 were analyzed by RT-qPCR using specific primers and expressed as fold change after normalization with β-actin level for each treatment. Mean ± SEM (n = 3), *, p < 0.05, **, p < 0.01, ***, p < 0.001, compared to untreated samples of day 1 or day 3, respectively. (C-F) Cells treated for 1 day (C and D) or 3 days (E and F) as above were lysed and analyzed by immunoblotting against Alox5 or β-actin (loading control). The representative image was shown from 3 different experiments. The relative amount of Alox5 was normalized to the amount of β-actin and expressed as % of untreated control at each day and quantification was shown as Mean ± SEM (n = 3), **, p < 0.01, ***, p < 0.001, compared to untreated samples of Day 1 and Day 3, respectively.