mouse anti enterovirus 71 antibodies  (Bio-Rad)

Journal: Scientific Reports

Article Title: Host neuronal PRSS3 interacts with enterovirus A71 3A protein and its role in viral replication

doi: 10.1038/s41598-022-17272-2

Figure Lengend Snippet: Determination of relative fold-changes of EV-A71 RNA copy numbers by qRT-PCR in response to overexpression of the neuron-derived PRSS3 in pLVX::FL-PRSS3-Myc -transfected HEK293T cells. ( a ) Cytotoxicity of the FL-PRSS3-Myc to HEK293T cells was determined by SRB assay. ( b ) The relative fold-changes of EV-A71 RNA copy numbers in response to overexpression of the FL-PRSS3-Myc. The pLVX::FL-PRSS3-Myc -transfected HEK293T cells infected with EV-A71 were designated as EV-A71-infected pLVX::FL-PRSS3-Myc -transfected cells. Mock transfected and pLVX- transfected HEK293T cells infected with EV-A71 served as infection control and empty vector control, respectively. ( c ) Western blot analysis of EV-A71 protein synthesis upon overexpression of the FL-PRSS3-Myc when compared with the control groups. The EV-A71 proteins detected by anti-EV-A71 antibody are VP0 (MW = 36 kDa), pre-cleavage product of VP2, and VP2 (MW = 28 kDa). Human β-actin (MW = 42 kDa) was used for normalization of protein loading. Data were expressed by mean ± SD from three independent experiments in triplicate measurements (N = 9). ( d ) The titers of virus released from cells overexpressed PRSS3, pLVX-Puro -transfected- and untransfected cells were determined by CCID50 method. The data was derived from three independent experiments. Statistical analysis was done by One-way ANOVA with post-hoc tests. Levels of statistically significant difference at p < 0.05 and p < 0.001 were indicated by * and ***, respectively. The original blots of ( c ) were shown in Supplementary Fig. .

Article Snippet: Mouse monoclonal anti-cMyc antibody and mouse anti-enterovirus 71 VP2 antibody were from Bio-Rad (USA).

Techniques: Quantitative RT-PCR, Over Expression, Derivative Assay, Transfection, Sulforhodamine B Assay, Infection, Control, Plasmid Preparation, Western Blot, Virus

Journal: Scientific Reports

Article Title: Host neuronal PRSS3 interacts with enterovirus A71 3A protein and its role in viral replication

doi: 10.1038/s41598-022-17272-2

Figure Lengend Snippet: Determination of relative fold-changes of EV-A71 RNA copy numbers by qRT-PCR in response to the knockdown of PRSS3 . ( a ) Fold-changes of gene expression of PRSS3 and ( b ) GAPDH in the siRNAs-transfected non-infected cells were relative to the mock transfected non-infected cells. ( c ) Relative fold-changes of EV-A71 RNA copy numbers in siPRSS3 -transfected HEK293T cells infected with EV-A71 when compared with siGAPDH- transfected- and untransfected infected cells. The qRT-PCR data were expressed by mean ± SD from three independent experiments in triplicate measurements (N = 9). ( d ) Western blot analysis of EV-A71 protein synthesis upon siRNA-mediated gene silencing of PRSS3 when compared with the control groups . The EV-A71 proteins detected by anti-EV-A71 antibody are VP0 (MW = 36 kDa), pre-cleavage product of VP2, and VP2 (MW = 28 kDa). Human β-actin (MW = 42 kDa) was used for normalization of protein loading. ( e ) The titers of virus released from PRSS3 -silenced cells, si - GAPDH transfected- and untransfected cells were determined by CCID50 method. The data was derived from triplicate experiments. Statistical analysis was done by One-way ANOVA with post- hoc tests. Levels of statistically significant difference at p < 0.05 and p < 0.001 were indicated by * and ***, respectively. The original blots of ( d ) were shown in Supplementary Fig. .

Article Snippet: Mouse monoclonal anti-cMyc antibody and mouse anti-enterovirus 71 VP2 antibody were from Bio-Rad (USA).

Techniques: Quantitative RT-PCR, Knockdown, Gene Expression, Transfection, Infection, Western Blot, Control, Virus, Derivative Assay