siRNA-mediated neddylation inhibition inhibits LPS-induced TNF-α production in JAWSII cells. (A) qPCR analysis of SAG expression in JAWSII cells transfected with indicated siRNA (3 μg). Scramble transfection performed as control. (B) qPCR analysis of TNF-α expression in JAWSII cells transfected with indicated siRNA and stimulated with LPS (0.5 µg/mL). Scramble transfection performed as control. One representative experiment of 3 is shown. (C-E) JAWSII cells transfected with siRNA for βTrCP, Cul5, or scramble as described in Materials and methods. Expression of βTrCP (C) and Cul5 (D) mRNA transcripts in JAWSII cells receiving indicated siRNA-mediated knockdown. (E) Expression of TNF-α mRNA transcripts in cells transfected with indicated siRNA and cultured in the presence or absence of LPS (0.5 µg/mL) for 4 hours. *P < .05; **P < .01; ***P < .0001.