Journal: Cells

Article Title: NXN Gene Epigenetic Changes in an Adult Neurogenesis Model of Alzheimer’s Disease

doi: 10.3390/cells11071069

Figure Lengend Snippet: Effect of Aβ peptide 1–42 addition on the NXN gene during the differentiation period. mRNA expression relative to the geometric mean of ACTB and GAPDH housekeeping genes expression ( A ). DNA methylation level in the extended mapping amplicon ( B ) were determined for the controls and Aβ peptide-treated neural progenitor cells on days 9, 19, and 29. Vertical lines represent the SEM. * p -value < 0.05.

Article Snippet: Briefly, 0.30 × 10 6 NPCs were seeded onto a CellMatrix Basement Membrane Gel (ATCC ® ACS3035TM) coated 12-well plate and incubated in NPC expansion medium: complete growth medium including DMEM/F-12 (Gibco, Fisher Scientific, Waltham, MA, USA), supplemented with the Growth Kit for Neural Progenitor Cell Expansion (ATCC ® ACS3003) and then maintained in a humidified incubator (5% CO 2 , 37 °C).

Techniques: Expressing, DNA Methylation Assay, Amplification

Journal: Scientific Reports

Article Title: Plasmid DNA gene therapy of the Niemann-Pick C1 mouse with transferrin receptor-targeted Trojan horse liposomes

doi: 10.1038/s41598-020-70290-w

Figure Lengend Snippet: Terminal organ weights (mg wet tissue) and body weights (grams) in female and male NPC1 −/− mice treated with either vehicle or THLs.

Article Snippet: A human NPC1 expression plasmid (#SC120010), under the influence of the CMV promoter, is designated pCMV-NPC1, and was purchased from Origene (Rockville, MD).

Techniques:

Journal: Scientific Reports

Article Title: Plasmid DNA gene therapy of the Niemann-Pick C1 mouse with transferrin receptor-targeted Trojan horse liposomes

doi: 10.1038/s41598-020-70290-w

Figure Lengend Snippet: H&E stains of cerebral cortex ( A – C ), spleen ( D – F ), and liver ( G – I ) from NPC1 −/− mice or control mice at euthanasia. Panels ( A , D , G ) are NPC1 −/− mice treated with vehicle; panels ( B , E , H ) are NPC −/− mice treated with TfRMAb/pPDGFB-NPC1 THLs. Panels ( C , F , I ) are control mice. Arrows in panels ( A , D , G ) point to cholesterol-laden cells in brain, spleen, and liver, respectively. Brain sections are through cortical layers I–IV. Magnification is the same for all panels and the panel A magnification bar = 100 microns.

Article Snippet: A human NPC1 expression plasmid (#SC120010), under the influence of the CMV promoter, is designated pCMV-NPC1, and was purchased from Origene (Rockville, MD).

Techniques: Control

Journal: Scientific Reports

Article Title: Plasmid DNA gene therapy of the Niemann-Pick C1 mouse with transferrin receptor-targeted Trojan horse liposomes

doi: 10.1038/s41598-020-70290-w

Figure Lengend Snippet: Electron microscopy of cerebral cortex of vehicle ( A , B ) or THL ( C , D ) treated NPC1 −/− mouse at euthanasia. Multi-lamellated vacuoles of varying size are present in the cytoplasm of brain cells. Magnification in panels ( A ) and ( C ) is the same and magnification bar in panel ( A ) is 1,000 nm; magnification in panels ( B ) and D is the same and magnification bar in panel ( B ) is 1,000 nm.

Article Snippet: A human NPC1 expression plasmid (#SC120010), under the influence of the CMV promoter, is designated pCMV-NPC1, and was purchased from Origene (Rockville, MD).

Techniques: Electron Microscopy

Journal: Scientific Reports

Article Title: Plasmid DNA gene therapy of the Niemann-Pick C1 mouse with transferrin receptor-targeted Trojan horse liposomes

doi: 10.1038/s41598-020-70290-w

Figure Lengend Snippet: GFAP immunocytochemistry of thalamus of NPC1 −/− mouse treated with vehicle ( A ) or the TfRMAb/pPDGFB-NPC1 THLs ( B ). The GFAP immunocytochemistry of thalamus from a control mouse is shown in ( C ). Sections are counter-stained with hematoxylin. Magnification of all 3 panels is the same and the panel ( A ) magnification bar = 100 microns.

Article Snippet: A human NPC1 expression plasmid (#SC120010), under the influence of the CMV promoter, is designated pCMV-NPC1, and was purchased from Origene (Rockville, MD).

Techniques: Immunocytochemistry, Control, Staining

Journal: Scientific Reports

Article Title: Plasmid DNA gene therapy of the Niemann-Pick C1 mouse with transferrin receptor-targeted Trojan horse liposomes

doi: 10.1038/s41598-020-70290-w

Figure Lengend Snippet: Organ concentrations (pg plasmid DNA per mg wet tissue) in female and male NPC1 −/− mice at 4 days following the last IV injection of either vehicle or THLs.

Article Snippet: A human NPC1 expression plasmid (#SC120010), under the influence of the CMV promoter, is designated pCMV-NPC1, and was purchased from Origene (Rockville, MD).

Techniques: Plasmid Preparation, IV Injection

Journal: Scientific Reports

Article Title: Plasmid DNA gene therapy of the Niemann-Pick C1 mouse with transferrin receptor-targeted Trojan horse liposomes

doi: 10.1038/s41598-020-70290-w

Figure Lengend Snippet: Organ enrichment of NPC1 mRNA, relative to GAPDH mRNA, in THL treated NPC1 −/− mice as compared to vehicle treated NPC1 −/− mice.

Article Snippet: A human NPC1 expression plasmid (#SC120010), under the influence of the CMV promoter, is designated pCMV-NPC1, and was purchased from Origene (Rockville, MD).

Techniques:

Journal: Scientific Reports

Article Title: Plasmid DNA gene therapy of the Niemann-Pick C1 mouse with transferrin receptor-targeted Trojan horse liposomes

doi: 10.1038/s41598-020-70290-w

Figure Lengend Snippet: NPC1 Western blot of the spleen of 3 vehicle treated NPC −/− mice and 3 THL treated NPC −/− mice. The spleens of the THL treated mice express the 200 kDa immunoreactive NPC1 protein.

Article Snippet: A human NPC1 expression plasmid (#SC120010), under the influence of the CMV promoter, is designated pCMV-NPC1, and was purchased from Origene (Rockville, MD).

Techniques: Western Blot

Journal: eLife

Article Title: Aβ-driven nuclear pore complex dysfunction alters activation of necroptosis proteins in a mouse model of Alzheimer’s disease

doi: 10.7554/eLife.92069

Figure Lengend Snippet: ( A ) Zeiss AiryScan confocal images of primary hippocampal neurons (DIV 14) labeled with pan-NPC antibody RL1 (magenta) and NUP98 (green) after linear deconvolution. A select region of the nuclear membrane (white box) is magnified (right-most three panels). Scale bar at 2 µm (original) and 0.5 µm (magnified). White arrowheads show colocalization of signals (white). ( B ) IMARIS 3D reconstruction and isometric view of a nucleus and the gated selection of RL1-positive (magenta) and NUP98-positive (green) puncta on the nuclear membrane (blue). ( C ) Pearson’s correlation coefficient for N=17 nuclei between RL1 and NUP98 signals. For a more complete statistical output, please refer to .

Article Snippet: NPC (RL1; mouse) , Santa Cruz Biotechnology (sc-58815) , 1:1000 (ICC), 1:500 (IHC).

Techniques: Labeling, Membrane, Selection

Journal: eLife

Article Title: Aβ-driven nuclear pore complex dysfunction alters activation of necroptosis proteins in a mouse model of Alzheimer’s disease

doi: 10.7554/eLife.92069

Figure Lengend Snippet:

Article Snippet: NPC (RL1; mouse) , Santa Cruz Biotechnology (sc-58815) , 1:1000 (ICC), 1:500 (IHC).

Techniques:

Journal: Virology

Article Title: Chinese hamster ovary cell lines selected for resistance to ebolavirus glycoprotein mediated infection are defective for NPC1 expression.

doi: 10.1016/j.virol.2012.05.018

Figure Lengend Snippet: Fig. 6. Resistant cell lines have mutations in NPC1. (a) Schematic of NPC1 protein. Location of insertions in R1 (star) and R2 (circle), and deletion (triangle) and substitution (diamond) in L2 indicated in diagram at top and in sequence at bottom. (b) Expression of NPC in WT, M12, and resistant cells transduced with pLenti CMV NPC1 Hygro or pLenti CMV GFP Hygro. (c) Resistant cells stably expressing NPC1 or GFP were challenged with VSV EboGP mCherry and infection was quantitated by flow cytometry. Bars represent average of means from three separate experiments done in triplicate; errors bars represent standard error means.

Article Snippet: The rLuc gene was removed from VSV-XN2 rLuc by cleavage with MluI and NheI and replaced with the GP-KpnIjunction-XhoI-mCherry sequence. pLenti CMV NPC1 Hygro was constructed by removing the GFP gene from pLenti CMV GFP Hygro and inserting into its place the NPC1 cDNA (Origene). pSport6 Axl (clone ID 5205825) was from Open Biosystems. pcDNA3.1 furin and pcDNA3.1-ACE2 were previously described (Simmons et al., 2005; Wool-Lewis and Bates, 1999).

Techniques: Sequencing, Expressing, Transduction, Stable Transfection, Infection, Cytometry