Fig. 8: RBP4 binds TLR4 to activate TRAF6-mediated ubiquitination of PCV2 ORF1 in macrophages. | Communications Biology

Fig. 8: RBP4 binds TLR4 to activate TRAF6-mediated ubiquitination of PCV2 ORF1 in macrophages.

From: Retinol binding protein 4 restricts PCV2 replication via selective autophagy degradation of viral ORF1 protein

Fig. 8

Immunoblotting analysis of PCV2 Cap protein level (a) or qPCR analysis of PCV2 DNA copies (b, left) or TCID50 assay of viral titers (b, right) in RBP4-KO 3D4/21 cells transfected with empty vector (EV) or Flag-pRBP4 plasmid for 6 h, then cells were treated for 6 h with or without TLR4-specific inhibitor TAK242 (1 μM), followed by infection with PCV2 for the indicated periods or 48 h. Numbers (lower) indicate the grayscale analysis on the protein bands of Cap and RBP4. c Immunoprecipitation analysis of RBP4-KO 3D4/21 cells expressing PCV2 ORF1 and His-Ub-K63 together with or without Flag-RBP4 as indicated for 6 h. Cells were untreated or treated with TAK242 (1 μM) for 18 h before harvest. Data are representative of three independent experiments (a, c) or pooled from three independent experiments (b, mean ± SD). *p < 0.05, **p < 0.01 (Student’s t test).

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