Figure 2

PCV3 Cap colocalizes and interacts with NPM1. A, B NPM1 colocalization with PCV3 Cap in transfected cells. HEK293T (A) and PK-15 (B) cells were cotransfected with GFP-PCV3-Cap and mCherry-NPM1 for 24 h, and cells were fixed and then subjected to confocal microscopy analysis. Nuclei were stained with 4′, 6′-diamidino-2-phenylindole (DAPI) (A, B). Scale bar, 10 μm. C. PK-15 cells were transfected with empty vector and Flag-PCV3-Cap for 48 h. D, E HEK293T cells were cotransfected with Myc-PCV3-Cap and Flag-NPM1 for 48 h. The cell lysates were immunoprecipitated with Flag beads (C, D) or anti-Myc purified IgG (E). F Lysates of Flag-PCV3-Cap transfected 293T cells mixed with the GST, GST-NPM1 proteins were pulled down with glutathione S-transferase (GST) beads, and then subjected to GST pull-down assays, followed by immunoblotting using corresponding antibodies.