Journal: PLoS ONE

Article Title: Characteristics and Predictive Value of Blood Transcriptome Signature in Males with Autism Spectrum Disorders

doi: 10.1371/journal.pone.0049475

Figure Lengend Snippet: Quantitative RT-PCR validations of 12 differentially expressed genes.

Article Snippet: LBR , Hs01032700_m1 , 2.50 , 7.55278E-10 , 1.63 , 5.85338E-05.

Techniques: Quantitative RT-PCR

Journal: Cytotechnology

Article Title: Characterization of lamin B receptor of Sf9 cells and its fate during Autographa californica nucleopolyhedrovirus infection

doi: 10.1007/s10616-020-00380-0

Figure Lengend Snippet: Cloning, expression and analysis of Sf9 lbr gene. a Total RNAs were isolated from Sf9 cells and the first cDNA was obtained by reverse transcription PCR. The lbr Orf was amplified from cDNA mixture with lbr-specific primers. The PCR products were run on 1% agarose gel. The molecular size of lbr in bp is shown. b Protein samples isolated from Sf9 cells were resolved on 10% SDS-PAGE, and transferred to PVDF membrane. The LBR was detected with a rabbit anti-LBR antibody and the bound antibody was detected with HRP-labeled secondary antibody. The protein size is shown on the left. c Sf9 cells were transiently transfected with piz-lbr-mcherry plasmids. The nucleus was stained with DAPI (blue) at 48 h post-transfection (h p.t.). The sub-cellular distribution of LBR (red) was observed using confocal fluorescence microscopy. The solid arrow shows the localization of intact LBR at the nuclear rim. Bar, 10 µm. d The structural analysis of Sf9 LBR. The LBR has eight transmembrane helices with the length of 20–23 amino acids in the C terminal (I–VIII). One of Ser/Thr-Pro-X-X (S/TPXX) motifs was shown in the diagram. The representative phosphorylation sites for CDK5, PKA, PKB, or PKC were also shown in the diagram. e The IP assay. The lysates of Sf9 cells were immunoprecipitated with an anti-Dm0-like lamin antibody ADL67 or normal IgG antibody. The cell lysates and immunoprecipitated proteins were separated by SDS-PAGE and then transferred to PVDF membranes. The transferred proteins were probed with anti-LBR antibody or ADL67 antibody. (Color figure online)

Article Snippet: The supernatants were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and detected by western blotting with rabbit anti-LBR antibody (diluted 1:1000) and anti-rabbit second antibody conjugated to horseradish peroxidase (diluted 1:5000; Boster, Wuhan, China).

Techniques: Clone Assay, Expressing, Isolation, Reverse Transcription, Amplification, Agarose Gel Electrophoresis, SDS Page, Membrane, Labeling, Transfection, Staining, Fluorescence, Microscopy, Immunoprecipitation

Journal: Cytotechnology

Article Title: Characterization of lamin B receptor of Sf9 cells and its fate during Autographa californica nucleopolyhedrovirus infection

doi: 10.1007/s10616-020-00380-0

Figure Lengend Snippet: Changes of the amount of LBR expression levels after AcMNPV infection. a Detection of the amount of LBR during AcMNPV infection by western blotting. Sf9 cells were infected with vAcBac at an MOI of 5. At 24 and 48 h p.i., the cells were harvested and the protein samples were separated on a 10% polyacrylamide gel and probed by western blotting using anti-LBR or anti-actin antibodies. b Quantitative analysis of the amount of LBR during AcMNPV infection. The amounts of LBR were quantified by measuring of band densities using Image-Pro 5.0 software. Mean values with standard deviations (SD) of three independent experiments were shown. One-way ANOVA was performed to compare the difference among different groups (*p < 0.05; **p < 0.01).

Article Snippet: The supernatants were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and detected by western blotting with rabbit anti-LBR antibody (diluted 1:1000) and anti-rabbit second antibody conjugated to horseradish peroxidase (diluted 1:5000; Boster, Wuhan, China).

Techniques: Expressing, Infection, Western Blot, Software

Journal: Cytotechnology

Article Title: Characterization of lamin B receptor of Sf9 cells and its fate during Autographa californica nucleopolyhedrovirus infection

doi: 10.1007/s10616-020-00380-0

Figure Lengend Snippet: The effect of PKC inhibitor on the LBR or lamin stability and infectious BV production. a The effect of PKC inhibitor on the LBR stability. Sf9 cells were infected with vAcBac at an MOI of 5. After incubation with the virus, PKC inhibitor was added to the culture medium. At 48 h p.i., total proteins were isolated and subjected to SDS-PAGE, followed by western blotting analysis with anti-LBR antibody. b The effect of PKC inhibitor on the BV production. Sf9 cells were infected with vAcBac at an MOI of 5 in the absence or presence of PKC inhibitor. At 24 h p.i., the supernatants were harvested and virus titers were calculated by end point dilution assay. Data are presented as mean ± standard deviations (SD) of triplicates. One-way ANOVA was performed to compare the differences between the two groups (**p < 0.01). c The effect of PKC inhibitor on the lamin stability. Sf9 cells were infected with vAcBac at an MOI of 5. After incubation with the virus, PKC inhibitor was added to the culture medium. At 48 h p.i., total proteins were resolved by SDS-PAGE and subjected to western blotting analysis using anti-Dm0-like lamin antibody.

Article Snippet: The supernatants were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and detected by western blotting with rabbit anti-LBR antibody (diluted 1:1000) and anti-rabbit second antibody conjugated to horseradish peroxidase (diluted 1:5000; Boster, Wuhan, China).

Techniques: Infection, Incubation, Virus, Isolation, SDS Page, Western Blot, End-point Dilution Assay