Skip to main content
Figure 4 | Journal of Neuroinflammation

Figure 4

From: Neuronal RING finger protein 11 (RNF11) regulates canonical NF-κB signaling

Figure 4

Dynamic associations of RNF11 with both A20 and Itch in primary neurons. N2A cells transduced with V5-RNF11 lentivirus (N2A V5-RNF11) and transfected with FLAG-A20 were harvested for immunoprecipitation (IP) with V5 antibody (A) or harvested for IP with FLAG antibody (B). Proteins were resolved by SDS-PAGE and immunoblotted with anti-A20 and RNF11. In parallel, pull-down assays with V5 antibody (C) or with Itch antibody (D) from N2A V5-RNF11 cell lysates were resolved by SDS-PAGE. Immunoprecipitates and lysates were immunoblotted with anti-Itch and RNF11. (E) and (H) Murine primary cortical neurons were stimulated with 10 ng/ml TNF-α for 0 or 30 minutes and harvested for IP with RNF11 antibody. Control IP experiments were performed with antibody omitted. Proteins were resolved by SDS-PAGE and immunoblotted with anti-A20, Itch, RNF11 and actin. (F), (G), (I) and (J) ImageJ software was used to quantify the densitometry of the immunoprecipitated bands relative to the 0-minutes time point. Each input sample’s immunoreactivity was used as a loading control. All IPs are representative of at least three independent experiments. * P < 0.05.

Back to article page