Journal: Heliyon

Article Title: Generation of rescued Japanese encephalitis virus genotype 1 from infectious full-size clone using reverse genetics

doi: 10.1016/j.heliyon.2024.e33142

Figure Lengend Snippet: Expression of JEV E and NS1 proteins. Vero cells were infected with rJEV-GI or-K05GS at an MOI of 0.1, and the cell culture supernatant and lysates were harvested at 5 dpi. (A) SDS-PAGE followed by Coomassie Brilliant Blue G-250 staining (B) Western blot analysis with specific antibodies against JEV E and NS1 protein. M, protein size marker; 1: mock-infected cell lysate; 2: rJEV-GI-infected cell lysate; 3: K05GS-infected cell lysate; 4: mock-infected cell supernatant; 5: rJEV-GI-infected cell supernatant; 6: K05GS-infected cell supernatant. Asterisks indicate a possible cleavage fragment of the JEV NS1 protein (50 kDa) following the instructions from Abcam (ab41651). This experiment was repeated twice, and no significant differences were found. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Anti -JEV envelope (E) antibody (GeneTex; 1:1000 dilution) and anti-JEV non-structural protein 1 (NS1) antibody (Abcam; 1:1000 dilution) were reacted with the membrane.

Techniques: Expressing, Infection, Cell Culture, SDS Page, Staining, Western Blot, Marker

Journal: International Journal of Molecular Sciences

Article Title: Lysophosphatidylcholines Promote Influenza Virus Reproduction through the MAPK/JNK Pathway in PMA-Differentiated THP-1 Macrophages

doi: 10.3390/ijms25126538

Figure Lengend Snippet: Increase in viral RNA and protein levels by LPCs. ( A ) Relative expression levels of IAV genes in THP-1 cells treated with different dosages of LPCs. Color boxes indicated control (□), IAV (■) and IAV with LPC (▒). ( B ) The protein level of IAV proteins in THP-1 cells treated with LPCs. RNA levels were determined using RT-qPCR, and protein expression was detected using Western blotting. Data represent the means ± SDs of three independent experiments. Statistical significance was assessed using an unpaired Student t -test. *** p < 0.001, ** p < 0.005, * p < 0.05 compared with the values obtained for cells infected with IAV. M2: matrix-2; M1: matrix-1; PA: polymerase acidic protein; PB1: polymerase basic 1; NP: nucleoprotein; NS1: non-structural protein; PB2: polymerase basic 2; HA: hemagglutinin; NA: neuraminidase.

Article Snippet: Antibodies against the influenza virus proteins polymerase acidic (PA), nucleoprotein (NP), non-structural protein 1 (NS1), matrix-1 (M1), polymerase basic 1 (PB1), and PB2 were obtained from GeneTex, Inc. (Irvine, CA, USA), and antibodies for JNK, p-JNK, AKT, and p-AKT were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot, Infection

Journal: International Journal of Molecular Sciences

Article Title: Lysophosphatidylcholines Promote Influenza Virus Reproduction through the MAPK/JNK Pathway in PMA-Differentiated THP-1 Macrophages

doi: 10.3390/ijms25126538

Figure Lengend Snippet: Differential expression of viral RNAs according to the type of LPCs. The relative expression levels of IAV genes were determined using RT-qPCR. Data represent the means ± SDs of three independent experiments. Statistical significance was assessed using an unpaired Student t -test. *** p < 0.001, ** p < 0.005, * p < 0.05 compared with the values obtained for cells infected with IAV. Color boxes indicated control (□), IAV (■) and IAV with LPCs (▒). M2: matrix-2; M1: matrix-1; PA: polymerase acidic protein; PB1: polymerase basic 1; NP: nucleoprotein; NS1: non-structural protein; PB2: polymerase basic 2; HA: hemagglutinin; NA: neuraminidase.

Article Snippet: Antibodies against the influenza virus proteins polymerase acidic (PA), nucleoprotein (NP), non-structural protein 1 (NS1), matrix-1 (M1), polymerase basic 1 (PB1), and PB2 were obtained from GeneTex, Inc. (Irvine, CA, USA), and antibodies for JNK, p-JNK, AKT, and p-AKT were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Quantitative Proteomics, Expressing, Quantitative RT-PCR, Infection, Control

Journal: International Journal of Molecular Sciences

Article Title: Lysophosphatidylcholines Promote Influenza Virus Reproduction through the MAPK/JNK Pathway in PMA-Differentiated THP-1 Macrophages

doi: 10.3390/ijms25126538

Figure Lengend Snippet: Attenuation of viral gene expression by MAP kinase inhibitor (SB203580), JNK inhibitor, and PI3K inhibitor (LY294002) in LPC-treated THP-1 cells. Relative expression levels of IAV genes were determined using RT-qPCR. Data represent the means ± SDs of three independent experiments. Statistical significance was assessed using an unpaired Student t -test. *** p < 0.001 compared with values obtained for cells infected with IAV, # p < 0.05 compared with values obtained for cells infected with IAV and treated with LPC. Color boxes indicated control (□), IAV (■) and IAV with LPC and inhibitors (▒). M2: matrix-2; M1: matrix-1; PA: polymerase acidic protein; PB1: polymerase basic 1; NP: nucleoprotein; NS1: non-structural protein; PB2: polymerase basic 2; HA: hemagglutinin; NA: neuraminidase.

Article Snippet: Antibodies against the influenza virus proteins polymerase acidic (PA), nucleoprotein (NP), non-structural protein 1 (NS1), matrix-1 (M1), polymerase basic 1 (PB1), and PB2 were obtained from GeneTex, Inc. (Irvine, CA, USA), and antibodies for JNK, p-JNK, AKT, and p-AKT were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Gene Expression, Expressing, Quantitative RT-PCR, Infection, Control