Fig. 3: RBP4 inhibits replication of PCV2 in vitro and in vivo. | Communications Biology

Fig. 3: RBP4 inhibits replication of PCV2 in vitro and in vivo.

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

Fig. 3

Immunoblotting analysis of PCV2 Cap protein levels (a) or TCID50 assay of viral titers (b) in 3D4/21 cells (left panel) or PK-15 (right panel) cells transfected with empty vector or Flag-RBP4 plasmid for 12 h followed by PCV2 infection for the indicated periods or 48 h. Immunoblotting analysis of PCV2 Cap protein levels (c) or TCID50 assay of viral titers (d) in wild-type (WT) and RBP4-deficient (RBP4-KO) 3D4/21 cells (left panel), WT and RBP4-KO PK-15 cells (right panel) infected with PCV2 for indicated periods or 48 h. Immunoblotting analysis of PCV2 Cap protein levels (e) or TCID50 assay of viral titers (f) in RBP4-KO 3D4/21 cells (left panel) and RBP4-KO PK-15 cells (right panel) infected with PCV2 for the indicated periods or 48 h. After 6 h of PCV2 infection, cells were treated with or without recombinant porcine RBP4 (rpRBP4, 30 μg/mL). g Immunoblotting analysis of PCV2 Cap protein levels (left panel) or TCID50 assay of viral titers (right panel) in primary PAMs infected with PCV2 for the indicated periods or 48 h. After 6 h of PCV2 infection, the cells were treated with or without recombinant porcine RBP4 (rpRBP4, 30 μg/mL). h Immunoblotting analysis of PCV2 Cap protein levels (left panel) or TCID50 assay of viral titers (right panel) in WT or RBP4-KO BMDMs infected with PCV2 for the indicated periods or 48 h. i Hematoxylin and eosin (H&E) staining of lung (left) and liver (right) sections from mice infected with PCV2 (5 × 105 pfu/mouse) for 7 days. Scale bar, 20 μm. Original magnification, ×40. j Histological scores of lung and liver from mice infected by PCV2 as in (i). Each symbol represents an individual mouse (n = 6/group). k Viral titers in the lung (left) and liver (right) from WT and RBP4-KO mice (n = 6/group) infected with PCV2 in (i). Data are pooled from three independent experiments (b, d, f and g, h right, j, k, mean ± SD) or representative of three independent experiments (a, c, e and g, h left, i). *p < 0.05, **p < 0.01 (Student’s t test).

Back to article page