Journal: NPJ Biofilms and Microbiomes

Article Title: Nasal symbiont Staphylococcus epidermidis restricts the cellular entry of influenza virus into the nasal epithelium

doi: 10.1038/s41522-022-00290-3

Figure Lengend Snippet: NHNE cells from five healthy volunteers were inoculated with human nasal S. epidermidis at an MOI of 0.25. A FemA mRNA levels, normalized to cellular GAPDH transcript levels, were monitored by real-time PCR. B S. epidermidis CFUs were determined at 1 dpi in the supernatant of S. epidermidis -inoculated NHNE cells. C Dot plot visualization of enriched GO terms in S. epidermidis– inoculated NHNE cells at 1 dpi. D Scatterplots indicating enriched genes correlated with serine-type peptidase inhibitors. E Expressions of Serpine1 and Serpine2 from scRNA-seq of NHNE cells. Dot size represents the proportion of Serpines RNA expression within the respective cell type expressing the gene, and dot color represent the average Serpines RNA expression level in the particular cell type. F Serpine1 and Serpine2 mRNA levels, normalized to cellular GAPDH transcript levels, were monitored by real-time PCR (blue bar: Serpine1; red bar: Serpine2). G The intracellular protein level of Serpine1 was measured in the cell lysate of S. epidermidis– inoculated NHNE cells using western blot analysis. H The secreted protein level of Serpine1 was measured in the supernatant of S. epidermidis -inoculated NHNE cells using ELISA. I Heatmap depicting the expression levels of genes related with serine-type peptidase inhibitor activity differentially expressed in S. epidermidis -inoculated NHNE cells depending on the cellular subset. J Serpine1 mRNA level, normalized to cellular GAPDH transcript levels, was monitored by real-time PCR over 1 day after IAV infection following S. epidermidis inoculation. K , L The neutralizing antibody for Serpine1 was administered to NHNE cells one hour before S. epidermidis inoculation, and then the cells were infected with IAV for 1 day. IAV mRNA and NPs of IAV were compared in IAV-infected NHNE cells after S. epidermidis inoculation depending upon the neutralization of Serpine1. Results are presented as mean ± SD values from five independent experiments. * p < 0.05 vs. mock-infected NHNE cells.

Article Snippet: In addition, a human Serpine1 antibody (molecular weight 45 kDa, primary antibody 1:500) was purchased from Abcam (Waltham, MA, USA, cat #ab66705) and mouse Serpine1 blocking peptide from MyBioSource (San Diego, CA, USA, cat #MBS8309331).

Techniques: Real-time Polymerase Chain Reaction, RNA Expression, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Activity Assay, Infection, Neutralization

Journal: NPJ Biofilms and Microbiomes

Article Title: Nasal symbiont Staphylococcus epidermidis restricts the cellular entry of influenza virus into the nasal epithelium

doi: 10.1038/s41522-022-00290-3

Figure Lengend Snippet: Differential expression gene (DEG) linked with serine-type peptidase inhibitors in S. epidermidis -inoculated NHNE cells.

Article Snippet: In addition, a human Serpine1 antibody (molecular weight 45 kDa, primary antibody 1:500) was purchased from Abcam (Waltham, MA, USA, cat #ab66705) and mouse Serpine1 blocking peptide from MyBioSource (San Diego, CA, USA, cat #MBS8309331).

Techniques: Quantitative Proteomics

Journal: NPJ Biofilms and Microbiomes

Article Title: Nasal symbiont Staphylococcus epidermidis restricts the cellular entry of influenza virus into the nasal epithelium

doi: 10.1038/s41522-022-00290-3

Figure Lengend Snippet: A Scatterplots indicating enriched genes related with serine-type peptidase. B PPI network of DEGs linked with serine-type peptidase indicates the protein – protein interaction with Serpine1 and Serpine2 (PP protein – protein). C Heatmap depicting the expression levels of genes related with serine-type peptidase inhibitor activity differentially expressed in S. epidermidis– inoculated NHNE cells depending on a cellular subset. D Serpine1 mRNA and E protein levels were monitored by real-time PCR and western blot analysis following S. epidermidis inoculation at 1 dpi with transfection with cont shRNA and uPA shRNA. F Serpine1 (blue bar) and PLAU mRNAs (red bar) were compared in NHNE cells after S. epidermidis inoculation. G ELISA results showed the secreted protein levels of Serpine1 (blue bar) and uPA (red bar) in S. epidermidis -inoculated NHNE cells. H The intracellular protein levels of HAT and Serpine1 were measured in the cell lysate of S. epidermidis– inoculated NHNE cells at 4, 8, and 24 h using western blot analysis. I The mRNA levels of proteases, such as TMPRSS2, TMPRSS11E, TMPRSS11F, and KLK5, which were targeted by Serpine1, were measured using the cell lysate of S. epidermidis -inoculated NHNE cells at 4, 8, and 24 h. Western blot results are representative of five independent experiments. Real-time PCR and plaque assay results are presented as mean ± SD values from five independent experiments. * p < 0.05 vs. mock-infected NHNE cells.

Article Snippet: In addition, a human Serpine1 antibody (molecular weight 45 kDa, primary antibody 1:500) was purchased from Abcam (Waltham, MA, USA, cat #ab66705) and mouse Serpine1 blocking peptide from MyBioSource (San Diego, CA, USA, cat #MBS8309331).

Techniques: Expressing, Activity Assay, Real-time Polymerase Chain Reaction, Western Blot, Transfection, shRNA, Enzyme-linked Immunosorbent Assay, Plaque Assay, Infection