Figure 3.
Figure 3. MyD88dn inhibits LPS-induced TNFα production in mouse RAW cells but not in perinoneal macrophages. RAW cells and murine peritoneal macrophages were left uninfected (white bars) or infected for 2 hours with adenoviruses overexpressing β-galactosidase (gray bars), MyD88wt (cross-hatched bars), or MyD88dn (black bars) in serum-free medium, and then cultured in complete medium for another day. (A) Cells were lysed and cytosolic extracts collected and examined for the expression of MyD88. RAW cells (B) and peritoneal macrophages (C) were treated with 1 μg/mL LPS for an additional 20 hours, and supernatants were assayed by ELISA for TNFα. Mean cytokine production (± SD) of triplicate cultures is shown and is representative of 3 independent experiments. Peritoneal macrophages were also treated with 1 μg/mL LPS or 20 ng/mL IL-1 for 45 minutes and nuclear extracts were collected and assayed for NF-κB activation by EMSA (D).

MyD88dn inhibits LPS-induced TNFα production in mouse RAW cells but not in perinoneal macrophages. RAW cells and murine peritoneal macrophages were left uninfected (white bars) or infected for 2 hours with adenoviruses overexpressing β-galactosidase (gray bars), MyD88wt (cross-hatched bars), or MyD88dn (black bars) in serum-free medium, and then cultured in complete medium for another day. (A) Cells were lysed and cytosolic extracts collected and examined for the expression of MyD88. RAW cells (B) and peritoneal macrophages (C) were treated with 1 μg/mL LPS for an additional 20 hours, and supernatants were assayed by ELISA for TNFα. Mean cytokine production (± SD) of triplicate cultures is shown and is representative of 3 independent experiments. Peritoneal macrophages were also treated with 1 μg/mL LPS or 20 ng/mL IL-1 for 45 minutes and nuclear extracts were collected and assayed for NF-κB activation by EMSA (D).

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