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Recombinant FliC and truncated FliC expressed from plasmid pORI induce TNF-α in a TLR5-dependent manner.
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posted on 2016-05-16, 08:02 authored by Jesse D. Deere, W. L. William Chang, Luis D. Castillo, Kim A. Schmidt, Hung T. Kieu, Nicholas Renzette, Timothy Kowalik, Stephen W. Barthold, Barbara L. Shacklett, Peter A. Barry, Ellen E. SpargerTNF-α ELISA of cell culture supernatants collected from TLR5-positive RAW424 (Blue) or its parental TLR5-negative RAW264.7 (Red) mouse macrophage cells. (A) RAW cells were treated with increasing concentrations of recombinant FliC. (B) Telomerized rhesus fibroblast cells were transiently transfected with plasmids containing full-length FliC (pORIfliC) or partially truncated FliCs (pORIfliCΔ196–378 or pORIfliCΔ141–398) and freeze thaw lysates were generated. Cell culture supernatants harvested from RAW cells treated with these lysates were assayed by an ELISA that measures TNF-α production. An average of two independent experiments and means with standard deviation are presented.
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mouse macrophage cell lineGagfusionWestern blot analysismacrophage cell linecodon optimized gag geneimmunodominant hypervariable domainvaccinerhesus macaques inoculatednovel RhCMV virusAIDSimmunodeficiency virustumor necrosis factorNonhuman Primate ModelSIV p 27 capsidOngoing animals studiesTLR 5 adjuvantproteinHIVtelomerized rhesus fibroblastTNF
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