AnapC5 binds to the IL-17 receptor and restricts IL-17-mediated signal transduction. A. AnapC5 associates with IL-17RA, IL-17RC and AnapC7.

<p>HEK293T cells were transfected with AnapC5 (tagged with HA) together with IL-17RA, IL-17RC or AnapC7 (tagged with Myc), as indicated. Lysates were subjected to IP with anti-Myc Abs and immunoblotted with anti-HA or anti-Myc. Arrows indicate identity of each Myc-tagged protein. Whole cell lysates (WCL) were verified for AnapC7 by staining with anti-HA. Migration of protein size markers is indicated. B. AnapC5 associates with IL-17RA through the inhibitory CBAD domain. HEK293T cells were transfected with AnapC5 and the indicated IL-17RA deletion constructs. Lysates were subjected to co-IP with anti-Myc and blotted for HA or Myc. Whole cell lysates (WCL) were verified for AnapC7 by staining with anti-Myc. Migration of protein size markers is indicated. C. AnapC5 associates with IL-17RC in the SEFIR domain. HEK293T cells were transfected with AnapC5 and the indicated IL-17RC deletion constructs. Lysates were subjected to co-IP with anti-Myc and blotted for HA or Myc. Whole cell lysates (WCL) were verified for AnapC7 by staining with anti-HA. Migration of protein size markers is indicated. D. Knockdown of AnapC5 enhances IL-17 signaling. ST2 cells were transfected with the indicated siRNAs, stimulated with IL-17 for 24 h, and IL-6 in culture supernatants assessed by ELISA. <i>n.s.,</i> not significant. *<i>p</i><0.05 by ANOVA and post-hoc Tukey’s test compared to unstimulated controls. ^‡<i>p</i><0.05 by Chi Square comparing experimental replicates. E. Efficient knockdown of AnapC5 and AnapC7. mRNA from the samples in panel D were assessed for AnapC5 and AnapC7 expression by qPCR. *<i>p</i><0.05 by ANOVA and post-hoc Tukey’s test compared to unstimulated controls. ^‡<i>p</i><0.05 by Chi Square comparing experimental replicates. Data are representative of at least 2 independent experiments.