Figure 1: TLR5 interacts with Nox1 and Nox4.
From: Flagellin-induced NADPH oxidase 4 activation is involved in atherosclerosis
(A) The direct interaction of the TLR5 with Nox1–5 C was estimated by yeast-two hybrid assay. Following the selection for Trp+ and His+ phenotype, Leu dependent growth and β-galactosidase activity were tested in an incubation medium (SD/galactose/Raffinose). (B) GST alone, GST-Nox1 (AA 217–550) and Nox4 (AA 248–578) were tested for their interactions with HEK293T cell lysates expressing Flag-tagged TLR5, and Flag-tagged TLR5 proteins were detected using an anti-Flag. Flag-CMV-TLR5 was transiently expressed in HEK293T cells either alone or together with pcDNA3.0-HA-Nox1C (AA 217–550) (C) or pcDNA3.0-HA-Nox4C (AA 250–573) (D), as indicated. After serum starvation, flagellin (100 ng/ml) was treated for 30 min. Cell lysates were then subjected to immunoprecipitation (IP) with antibodies to Flag, and the resulting precipitates were subjected to immunoblot analysis with antibodies to HA.
