Journal: Science Advances

Article Title: The nuclear import receptor importin-7 targets HPV from the Golgi to the nucleus to promote infection

doi: 10.1126/sciadv.adz6792

Figure Lengend Snippet: ( A ) HeLa S3 cells were uninfected or infected with HPV16.L2F (MOI, ~100). PLA (green) for IPO7 and FLAG (to detect L2) was performed at 24 hpi; nuclei were stained with DAPI (blue). Similar results were obtained in two independent experiments. Scale bar, 10 μm. ( B ) PLA fluorescence intensity per cell (>100 cells) as in (A), shown as individual values, means, and SDs, with statistical significance determined by one-way ANOVA. **** P < 0.0001. ( C ) Whole-cell extracts from uninfected or HPV16.L2F-infected (MOI, ~1) HeLa cells were collected at the indicated time points. A portion of the resulting extracts (input) was analyzed by immunoblotting for FLAG (to detect L2) and IPO7. The remaining extracts were subjected to IP with an anti-IPO7 antibody or control IgG and analyzed by immunoblotting for FLAG and IPO7. ( D ) HeLa cells were pretreated with 9 μM RO3306 or DMSO as a control for 24 hours and then uninfected or infected with HPV16.L2F (MOI, ~1), with RO3306 or DMSO maintained in the medium. At 24 hpi, cells were lysed and analyzed in (C). ( E ) HeLa cells treated with 10 nM Scr or a mixture of 5 nM COPA and 5 nM COPG1 ( COPA/G1 ) siRNA were uninfected or infected with HPV16.L2F PsV (MOI, ~1). At 24 hpi, cells were lysed, and a portion of the extracts (input) was immunoblotted for FLAG, IPO7, γ-COP, or β-actin (as a loading control). The remaining extracts were subjected to IP with an anti-IPO7 antibody or a control IgG and analyzed as in (C). ( F ) As in (E), except cells were transfected with 10 nM Scr or IPO7 siRNA #2 before infection. Samples were subjected to IP with an anti–γ-COP antibody or control IgG and analyzed as in (E).

Article Snippet: γ-COP , Rabbit , 12393-1-AP; Proteintech , WB and IP.

Techniques: Infection, Staining, Fluorescence, Western Blot, Control, Transfection

Journal: Veterinary Research

Article Title: Comprehensive multiomic analysis of extracellular vesicles from Mycoplasma bovis -infected bovine mammary epithelial cells identifies proteins and miRNAs that induce inflammatory responses in macrophages

doi: 10.1186/s13567-025-01626-5

Figure Lengend Snippet: Quantitative proteomic analysis of differentially expressed proteins in Ctrl-EVs and M. bovis NX2-EVs . A Venn diagram showing common and unique proteins in Ctrl-EVs and M. bovis NX2-EVs. B Statistics on the identification and quantification of proteins in Ctrl-EVs and M. bovis NX2-EVs. C Histogram of the subcellular localization of differentially expressed proteins between M. bovis NX2-EVs and Ctrl-EVs. D GO enrichment analysis of all significantly differentially expressed proteins; BP (biological process), CC (cellular component) and MF (molecular function) are indicated by green, blue and red, respectively. E KEGG enrichment analysis of all significantly differentially expressed proteins. F Volcano diagram of differentially expressed proteins between M. bovis NX2-EVs and Ctrl-EVs; red and blue dots indicate significantly upregulated and downregulated proteins, respectively, whereas grey dots represent proteins whose expression was not significantly different. G The abundance of the JCHAIN, JAG1, MAPRE1, ARCN1, APLP2, TSG101 and CD63 proteins was determined by western blotting. H Representative TEM of EVs labelled with immunogold using anti-JCHAIN, anti-JAG1, anti-MAPRE1, anti-ARCN1 and anti-TSG101 Antibodies. Scale bar: 100 nm.

Article Snippet: The grids were subsequently incubated for 2 h at 4 °C with appropriate dilutions of the primary antibodies against ARCN1 (1:1000, ProteinTech), MAPRE1 (1:1000, Affinity), JAG1 (1:1000, ProteinTech), TSG101 (1:1000, ProteinTech) and JCHAIN (1:1000, AtaGenix), and after repeating the washing step (3 min each time), the grid was floated for 2 h at room temperature on Anti-rabbit IgG, Anti-mouse IgG, and secondary antibody droplets containing 10-nm gold particles (AURION, Hatfield, PA).

Techniques: Expressing, Western Blot