Fig. 4

PRRSV infection induces increase of cyclinB1 expression. a Detection of cyclinB1 expression with western blot. MARC-145 cells mock-infected and 1 MOI PRRSV-infected were collected at 24 h and 48 h after PRRSV infection. CyclinB1 expression was detected with western blot using a specific antibody against cyclinB1. MARC-145 cells treated with 50 ng/mL nocodazole(Noco.) for 16 h served as a positive control (left), and expression levels were quantitatively analyzed and compared with GAPDH expression using Image J(https://imagej.nih.gov/ij/index.html) (right). ** indicates p < 0.01, *** indicates p < 0.001. b Detection of cyclinB1 expression and localization with IFA. Mock- and PRRSV-infected MARC-145 cells at 48 h post-infection were stained with an anti-cyclinB1 antibody, Phalloidin, and DAPI to determine cyclinB1(red), filamentous actin (F-actin) (green), and DNA (blue). Phalloidin(Phalloidin belongs to a class of toxins called phallotoxins. It functions by binding and stabilizing F-actin and effectively prevents the depolymerization of actin fibers. The properties of phalloidin make it a useful tool for investigating the distribution of F-actin in cells by labeling phalloidin with fluorescent analogs and using them to stain F-actin for light microscopy.) was used to show the outline of the cells. Then, the cells were visualized using Leica microsystems (Leica AF6000, Germany) (× 200)