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| Table 1: MCSs inhibit NF-kappa B p50 and p65 DNA binding in U937s. U937 cells were cultured with or without MSCs and followed by stimulation with LPS (5ng/ml) to mobilize nuclear translocation of NF-kappa B. Nuclear extracts were prepared using the Nuclear Extract Kit from Active Motif (Carlsbad, CA). p50 and p65 DNA binding was quantified by Trans-AM NF-kappa B DNA binding ELISA from Active Motif. NF-kappa B subunit p50 was significantly less active in nuclear extracts from U937s co-cultured with MSCs compared to nuclear extracts from U937s stimulated with PMA alone or U937s co-cultured with PECs as a control cell (0.63 ± 0.03 vs. 0.92 ± 0.14 and 0.63 ± 0.03 vs. 1.7 ± 0.3 respectively, p=0.05). DNA binding of the p65 subunit of NF-kappa B was also significantly inhibited in U937s cultured with MSCs versus U937s stimulated with PMA or U937s co-cultured with PECs as a control cell (0.34 ± 0.06 vs. 0.55 ± 0.01 and 0.34 ± 0.06 vs. 0.793 ± 0.07, respectively, p<0.05). |
