Cauliflower Mosaic Virus TAV, a Plant Virus Protein That Functions like Ribonuclease H1 and is Cytotoxic to Glioma Cells

Figure 2

TAV detection with western blot (WB), transmission electron microscopy (TEM), and confocal microscopy. (a) Western blot shows total proteins (w) and nuclear proteins (n) of EV and TAV samples, immunoblotted with anti-TAV polyclonal serum. Arrow indicates TAV band (62 kDa), present in TAV sample only, in both W and N. Since the TAV antibody is a rabbit polyclonal antisera, it showed also a nonspecific band, also in EV sample just above the TAV band. Full-length blots/gels are presented in Supplementary Figure 1. (b) Western blot shows total proteins (w) and cytosolic proteins (c) of each sample, immunoblotted with anti-TAV polyclonal serum. Arrow indicates the TAV band, present in the TAV sample only. Full-length blots/gels are presented in Supplementary Figure 2. (c–h) Immunogold electron microscopy showing localization of CaMV TAV in Hek293T (c, d, e) and turnip cells (f, g, h). Ultrathin sections were immunostained with anti-TAV and gold-labelled secondary antibody. EV (c) and noninfected turnip cells (f) were used as a negative reference. Gold particles are indicated by arrows at low magnification (d, g) and clearly visible at higher magnification (e, h). The bars represent 200 nm. The letter symbols stand for the following: N: nucleus; Nu: nucleolus; NM: nuclear membrane; M: mitochondria; L: lysosome; RE: endoplasmic reticulum; CW: cell wall; CH: chloroplast; V: vacuole; S: starch.