dyssey IR CLx system using the Odyssey imaging software 3.0 (Li-Cor Biosciences). Scan settings were high image quality, 169 m resolution, intensity 6.0 for the 700-channel, and 6.0 for the 800-channel with an offset of 4.0 mm. For signal quantification, antibody signals were analyzed as the average 800-channel integrated intensities from duplicate wells normalized to the 700-channel signal integrated intensity to correct for well-to-well variations in cell number. Results are expressed as percent inhibition of the NF-B responses (means standard errors of the mean) compared to vehicle-treated controls.
Active mitochondria with high membrane potential (Cm) accumulate the lipophilic cationic probe 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide (JC-1) in aggregates, which are red, whereas, in the mitochondria with low Cm (inactive), JC-1 stays in a monomeric, green form [38,39]. This renders the red:green ratio, a sensitive indicator of the mitochondrial Cm changes, and indicates cellular ROS production. The change in the mitochondrial membrane potential was measured in fullerene PTK/ZK derivative-treated human MC using JC-1 as previously described [40]. Cells (5×105/500 l) were incubated overnight with fullerene derivative as above, washed, loaded with 2 M JC-1 for 15 minutes, and activated as above.After cell stimulation, the green fluorescence (the monomeric JC-1) and red fluorescence (JC-1 aggregates) were measured using the FL-1 and FL-2 channels, respectively, with flow cytometry (Becton Dickinson, FACSCalibur, East Rutherford, New Jersey, USA). Mast cells were incubated overnight with fullerene derivatives, washed, and incubated for 30 minutes with 2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA, final concentration of 5 M). Next, cells were washed and resuspended in fresh media, placed in a cuvette and activated with IC as above for 5000 seconds. ROS fluorescence intensity was measured at 523 nm wavelength over a 12 minute time interval using spectrophotometry 
(Perkin Elmer, LS55 Luminescent Spectrometer, Waltham, Massachusetts, USA). All samples were measured in duplicate and performed at least three times.
To track the fate of fullerene derivatives in vivo an IRDye 800CW conjugated to a C70 fullerene using a protocol as described [41]. These dyes are used in conjunction with the Xenogen imaging system and have been widely used for bio-distribution studies [42]. The success of the conjugation and removal of free dye was verified using MALDI-MS (Bruker Corporation, Billerica, Massachusetts, USA) and absorption spectra. Live mice with or without full-blown disease (K/ BxN; day 14) were injected with various concentrations of the fullerene-dye conjugate and whole body images obtained over 24 hours.K/BxN 17764671 serum (125 l) was injected intraperitoneally (i.p.) on experimental days 0 and 1. Fullerene derivatives (40 g/100 l PBS) were injected i.p into C57BL/6 or mast cell-deficient Cremaster mice one day before the first serum injection and then every other day. Clinical scoring is detailed below. Ankle swelling i were measured using calipers along with the clinical indices as described [29]. In some experiments, serum was collected at day 14 and assayed for TNF- levels by ELISA (R&D systems, Minneapolis, Minnesota, USA). Mice were sacrificed, ankle sections removed, and sections scored as described below. Animal studies for the Cre-Master mice were approved by the Dana Farber Cancer Institute. To examine fullerene deri
