Journal: Viruses

Article Title: In Vitro Antiviral Properties of Two Recombinant Sendai Virus Vectors Encoding ORFV 011 and ORFV 059 Genes

doi: 10.3390/v18040462

Figure Lengend Snippet: HEK293T cells transfected with SeV-GFP, rSeV-GFP-B2L or rSeV-GFP-059. The transfection efficiency was determined by the presence of GFP-positive cells under fluorescence microscopy at 24, 96 and 144 h post-transfection (hpt). (Scale bar = 15 µm).

Article Snippet: ORFV 011 (1206 base pairs (bp)) and ORFV 059 (1029 bp) gene sequences were amplified from the ORFV strain NAV and cloned into the SeV-GFP plasmid by In-Fusion ® cloning technology, between Gaussia-Dura Luc and GFP genes (primers in ; In-Fusion HD Cloning Kit; Takara Bio USA, San Jose, CA, USA), generating recombinant plasmids rSeV-GFP-B2L and rSeV-GFP-059.

Techniques: Transfection, Fluorescence, Microscopy

Journal: Viruses

Article Title: In Vitro Antiviral Properties of Two Recombinant Sendai Virus Vectors Encoding ORFV 011 and ORFV 059 Genes

doi: 10.3390/v18040462

Figure Lengend Snippet: Relative mRNA expression of upregulated TLRs , RIG-I , MyD88 and IFN-β genes in OSF transduced with SeV-GFP, rSeV-GFP-B2L or rSeV-GFP-059 at 12 ( a ), 24 ( b ), 48 ( c ) and 72 ( d ) hours post-transduction (hpt). Genes with expression values >1 (broken line) were considered to be upregulated. Data represent the mean and standard deviation. Statistically significant differences between groups (* p < 0.05).

Article Snippet: ORFV 011 (1206 base pairs (bp)) and ORFV 059 (1029 bp) gene sequences were amplified from the ORFV strain NAV and cloned into the SeV-GFP plasmid by In-Fusion ® cloning technology, between Gaussia-Dura Luc and GFP genes (primers in ; In-Fusion HD Cloning Kit; Takara Bio USA, San Jose, CA, USA), generating recombinant plasmids rSeV-GFP-B2L and rSeV-GFP-059.

Techniques: Expressing, Transduction, Standard Deviation

Journal: Viruses

Article Title: In Vitro Antiviral Properties of Two Recombinant Sendai Virus Vectors Encoding ORFV 011 and ORFV 059 Genes

doi: 10.3390/v18040462

Figure Lengend Snippet: Relative mRNA expression of interferon-stimulated genes ( A3Z1 , OBST2 and SAMHD1 ) in OSF transduced with SeV-GFP, rSeV-GFP-B2L or rSeV-GFP-059 at 24 ( a ), 48 ( b ) and 72 ( c ) hours post-transduction (hpt). Dotted line corresponds to non-transduced OSF. Data shown are the mean and standard deviation. Statistically significant differences between groups (*** p < 0.001).

Article Snippet: ORFV 011 (1206 base pairs (bp)) and ORFV 059 (1029 bp) gene sequences were amplified from the ORFV strain NAV and cloned into the SeV-GFP plasmid by In-Fusion ® cloning technology, between Gaussia-Dura Luc and GFP genes (primers in ; In-Fusion HD Cloning Kit; Takara Bio USA, San Jose, CA, USA), generating recombinant plasmids rSeV-GFP-B2L and rSeV-GFP-059.

Techniques: Expressing, Transduction, Standard Deviation

Journal: Viruses

Article Title: In Vitro Antiviral Properties of Two Recombinant Sendai Virus Vectors Encoding ORFV 011 and ORFV 059 Genes

doi: 10.3390/v18040462

Figure Lengend Snippet: ORFV DNA quantification in KOP-R transduced with SeV-GFP, rSeV-GFP-B2L or rSeV-GFP-059 and infected (48 h post-transduction) with ORFV strain NAV. DNA were quantified at 24 h post-infection (hpi) and 5 days post-infection (dpi). Non-transduced KOP-R cells infected with ORFV strain NAV at the mentioned times served as positive controls (C+ (ORFV)). Data represent the mean and standard deviation. Statistically significant differences between groups (* p < 0.05).

Article Snippet: ORFV 011 (1206 base pairs (bp)) and ORFV 059 (1029 bp) gene sequences were amplified from the ORFV strain NAV and cloned into the SeV-GFP plasmid by In-Fusion ® cloning technology, between Gaussia-Dura Luc and GFP genes (primers in ; In-Fusion HD Cloning Kit; Takara Bio USA, San Jose, CA, USA), generating recombinant plasmids rSeV-GFP-B2L and rSeV-GFP-059.

Techniques: Transduction, Infection, Standard Deviation

Journal: Viruses

Article Title: In Vitro Antiviral Properties of Two Recombinant Sendai Virus Vectors Encoding ORFV 011 and ORFV 059 Genes

doi: 10.3390/v18040462

Figure Lengend Snippet: Relative mRNA expression of ORFV 045 in KOP-R transduced with SeV-GFP, rSeV-GFP-B2L or rSeV-GFP-059 and infected (48 h post-transduction) with ORFV strain NAV. Expression of ORFV 045 was quantified 24 and 48 h post-infection (hpi). Non-transduced KOP-R cells infected with ORFV strain NAV at the mentioned times served as positive controls (C+ (ORFV)). Data represent the mean and standard deviation.

Article Snippet: ORFV 011 (1206 base pairs (bp)) and ORFV 059 (1029 bp) gene sequences were amplified from the ORFV strain NAV and cloned into the SeV-GFP plasmid by In-Fusion ® cloning technology, between Gaussia-Dura Luc and GFP genes (primers in ; In-Fusion HD Cloning Kit; Takara Bio USA, San Jose, CA, USA), generating recombinant plasmids rSeV-GFP-B2L and rSeV-GFP-059.

Techniques: Expressing, Transduction, Infection, Standard Deviation

Journal: Viruses

Article Title: In Vitro Antiviral Properties of Two Recombinant Sendai Virus Vectors Encoding ORFV 011 and ORFV 059 Genes

doi: 10.3390/v18040462

Figure Lengend Snippet: Evaluation of the ORFV-specific cytopathic effect 5 days post-infection with ORFV strain NAV in non-transduced-KOP-R cells ( a ) and KOP-R cells transduced with SeV-GFP ( b ), rSeV-GFP-B2L ( c ) and rSeV-GFP-059 ( d ). The x, y values denote the calibrated pixel resolution in micrometers (µm) for the respective micrographs.

Article Snippet: ORFV 011 (1206 base pairs (bp)) and ORFV 059 (1029 bp) gene sequences were amplified from the ORFV strain NAV and cloned into the SeV-GFP plasmid by In-Fusion ® cloning technology, between Gaussia-Dura Luc and GFP genes (primers in ; In-Fusion HD Cloning Kit; Takara Bio USA, San Jose, CA, USA), generating recombinant plasmids rSeV-GFP-B2L and rSeV-GFP-059.

Techniques: Infection, Transduction

Journal: Viruses

Article Title: In Vitro Antiviral Properties of Two Recombinant Sendai Virus Vectors Encoding ORFV 011 and ORFV 059 Genes

doi: 10.3390/v18040462

Figure Lengend Snippet: SRLV DNA quantification in OSF incubated with supernatants from OSF transduced with SeV-GFP, rSeV-GFP-B2L or rSeV-GFP-059 and infected with SRLV strain EV1 after 24 h. Supernatants were collected at 24, 48 and 72 h post-transduction (hpt). Non-transduced OSF infected with EV1 at the mentioned times served as positive controls (C+ EV1). Data shown are the mean and standard deviation. Statistically significant differences between groups (** p < 0.01).

Article Snippet: ORFV 011 (1206 base pairs (bp)) and ORFV 059 (1029 bp) gene sequences were amplified from the ORFV strain NAV and cloned into the SeV-GFP plasmid by In-Fusion ® cloning technology, between Gaussia-Dura Luc and GFP genes (primers in ; In-Fusion HD Cloning Kit; Takara Bio USA, San Jose, CA, USA), generating recombinant plasmids rSeV-GFP-B2L and rSeV-GFP-059.

Techniques: Incubation, Transduction, Infection, Standard Deviation

Journal: bioRxiv

Article Title: Targeting Nitric Oxide Synthase 2 Reverses Learning Deficits in an Oligodendrocyte-Focused Model of Costello Syndrome

doi: 10.64898/2026.06.01.729333

Figure Lengend Snippet: A) Recombination strategy for tamoxifen (tmx; cyan)–mediated replacement of the HRas gene (grey, flanked by loxP sites) with HRasG12V (purple) in PlpCreERt2;FR-HRasG12V (pHRsG/+) mice. B) 2-month-old (2MO) mice treated with tmx are subjected to the complex wheel (CW) test, as well as histological and DTI-MRI analyses, 1-16 weeks later. C) Cell populations are analyzed in seven regions throughout the anterior-posterior (I-IV) and lateral “B” - central “C” axes of the corpus callosum (CC), unless otherwise disclosed. D) Immunostaining of coronal sections showing recombinant cells (GFP+, green arrowheads), OLs (GSTpi+, purple arrowheads), and GFP+GSTpi+ recombinant OLs (orange arrowheads) in the CC of WT and pHRsG/+ mice. E) Bar graph showing that the percentage (normalized to dapi) of recombinant cells in WT and pHRsG/+ mice is not significantly different (unpaired Student’s t test; males P= 0.14 and females P= 0.24). Genotype/sex color code and “n” per group in E, G : male WT (green) and pHRsG/+ (orange); female WT (black) and pHRsG/+ (red). F) Immunostaining picture indicating recombinant cells (green arrowheads), microglia (IBA1+, blue arrowheads), and OPCs (PDGFRa+, orange arrowheads) in the CC of WT and pHRsG/+ mice. G) The percentage of IBA+ cells in WT and pHRsG/+ mice (% of age/gender matched WTs) indicates significantly decreased # of microglia in male pHRsG/+ mice (unpaired Student’s t test; P= 0.047). Insets in D and F show the CC region in high magnification (dotted yellow square). Dapi was used to stain nuclei and normalize cell densities per arbitrary units (AU). Scale bar = 25μm. *P < 0.05.

Article Snippet: Floating sections were processed for immunodetection using antibodies for the reporter gene GFP (Nacalai Tesque, Kyoto, Japan), and the cell-type markers GSTpi (MBL Ltd, Tokio, Japan), CC1 (Calbiochem, San Diego, CA, USA), NG2 (Millipore, Burlington, MA, USA), PDGFRa (Rn’D systems, Minneapolis, MN), GFAP (Dako, Santa Clara, CA, USA), IBA1 (Wako, Osaka, Japan), and NeuN (Millipore, St. Louis, MO).

Techniques: Immunostaining, Recombinant, Staining

Journal: bioRxiv

Article Title: A neuropsychiatric disease-associated mutation in LRRC8B disrupts cellular Ca²⁺ signaling, mitochondrial function, and bioenergetics

doi: 10.64898/2026.04.16.718892

Figure Lengend Snippet: Volcano plot showing LC–MS/MS–based proteomic analysis of GFP pull-down samples from HEK293T cells expressing GFP-tagged wild-type (WT) or mutant LRRC8B. The x-axis represents log₂ fold change (mutant vs WT), and the y-axis shows −log₁₀ adjusted p-value. Proteins enriched in WT samples are shown on the left, while those enriched in mutant samples are shown on the right. Selected significantly enriched proteins are labeled. VDAC1 is identified as a WT-enriched interactor.

Article Snippet: The plasmid encoding GFP-tagged human LRRC8B (HG24935-ACG; Sino Biological Inc, USA).

Techniques: Liquid Chromatography with Mass Spectroscopy, Expressing, Mutagenesis, Labeling

Journal: bioRxiv

Article Title: A neuropsychiatric disease-associated mutation in LRRC8B disrupts cellular Ca²⁺ signaling, mitochondrial function, and bioenergetics

doi: 10.64898/2026.04.16.718892

Figure Lengend Snippet: Representative immunoblot images showing VDAC protein levels in total cell lysates and isolated mitochondrial fractions from cells transfected with GFP-tagged LRRC8B WT (W) or mutant Y380S (M) constructs. Actin (∼42 kDa) is included as a loading control for total lysates. Red boxes indicate the regions that were cropped and presented in the main figures. Molecular weight markers are shown where applicable.

Article Snippet: The plasmid encoding GFP-tagged human LRRC8B (HG24935-ACG; Sino Biological Inc, USA).

Techniques: Western Blot, Isolation, Transfection, Mutagenesis, Construct, Control, Molecular Weight

Journal: bioRxiv

Article Title: A neuropsychiatric disease-associated mutation in LRRC8B disrupts cellular Ca²⁺ signaling, mitochondrial function, and bioenergetics

doi: 10.64898/2026.04.16.718892

Figure Lengend Snippet: (A) Representative immunoblot images showing protein levels of GFP-tagged LRRC8B (WT and mutant) and VDAC in input lysates used for pull-down assays. (B–C) Immunoblot analysis of four independent immunoprecipitation (IP) experiments. Bands at ∼120 kDa confirm successful pull-down of GFP-tagged LRRC8B (WT and mutant) using an anti-GFP antibody. VDAC (∼32 kDa) bands indicate co-precipitation of VDAC with LRRC8B WT and mutant proteins. VDAC levels were normalized to the corresponding LRRC8B (WT or mutant) levels for quantification. Red boxes highlight the regions that were cropped and presented in the main figures. Molecular weight markers are shown where applicable.

Article Snippet: The plasmid encoding GFP-tagged human LRRC8B (HG24935-ACG; Sino Biological Inc, USA).

Techniques: Western Blot, Mutagenesis, Immunoprecipitation, Molecular Weight

Journal: Cancers

Article Title: SNAT1 (SLC38A1) Is Not the Main Glutamine Transporter in Melanoma, but Controls Metabolism via Glutamine-Dependent Activation of P62 (SQSTM1)/cMYC-Axis

doi: 10.3390/cancers18071068

Figure Lengend Snippet: Validation of cMYC as a downstream effector of SNAT1 regarding ATP synthesis and mitochondrial respiration. ( A ) Exemplary measurement of OCR of Mel Im conducting the ATP rate assay. Melanoma cells were transfected with ZsGreen1-cMYC/pLVX-Puromycin or pLenti-C-GFP, respectively. ( B ) Calculation of ATP generated via glycolysis and oxidative phosphorylation, total ATP and basal respiration of Mel Im and Mel Juso. ATP production rate and OCR were normalized to 10,000 cells and compared to siCtr. Values represent the mean ± SEM of at least three independent experiments. The treatment and respective control group were compared using the Student’s unpaired t -test. p -value < 0.05 was considered statistically significant (*).

Article Snippet: All of the following functional experiments were conducted after a total transfection period of 96 h. Transfection with ZsGreen1-cMYC/pLVX-Puromycin vector was performed using 0.5 μg plasmid DNA and Lipofectamine LTX Plus reagent (Invitrogen, ThermoFisher Scientific) for a transfection period of 48 h. Andre Catic kindly gifted the ZsGreen1-cMYC/pLVX-Puromycin vector (RRID:Addgene_180278). pLenti-C-GFP (OriGene, Rockville, MD, USA; RRID:Addgene_17448) served as a control plasmid in our experiments.

Techniques: Biomarker Discovery, Transfection, Generated, Phospho-proteomics, Control