fluorescent protein gfp reporter gene  (InvivoGen)

Journal: Frontiers in virology (Lausanne, Switzerland)

Article Title: RSV infection of humanized lung-only mice induces pathological changes resembling severe bronchiolitis and bronchopneumonia

doi: 10.3389/fviro.2024.1380030

Figure Lengend Snippet: (A) LoM were administered a single intramuscular dose of palivizumab 24 h prior to RSV A2-GFP exposure (15mg/kg or 30mg/kg) or 24 h post-RSV A2-GFP exposure (30mg/kg). Untreated RSV-GFP infected LoM served as a control. (B) GFP+ cells in human lung implants 4 days post-RSV exposure in untreated LoM (n=7 implants) and LoM administered palivizumab pre (15 mg/kg, n=6 implants and 30 mg/kg, n=3 implants) or post (n=4 implants) RSV exposure. (C) Ribavirin was administered to mice once daily (40 mg/kg, intraperitoneal) starting 24 h before or after RSV exposure. Untreated RSV-Luc infected LoM served as a control. (D) Bioluminescence signal (radiance [p sec–1 cm–2 sr–1] represented as total flux) in human lung implants just prior to RSV-exposure and 7 days post-RSV exposure in untreated LoM (white bars, n= 5 implants) and LoM administered ribavirin treatment initiated pre (orange bars, n=6 implants) or post (green bars, n= 6 implants) RSV exposure. The mean (horizontal line) and standard deviation (vertical line) are shown. Background luminescence measured pre-exposure is denoted by the dashed line. (E) Immunohistochemical staining for RSV antigen (brown) in human lung implants 7 days post RSV exposure in untreated LoM (left panel) and LoM administered ribavirin treatment-initiated pre (middle panel) or post (right panel) RSV exposure (scale bars, 100 um; n=3 implants analyzed). (B and D) Data are shown as mean ± SD. Results from treated groups were compared to untreated controls using a two-tailed Kruskal-Wallis test and P values were adjusted for multiple testing using the Benjamini, Krieger, Yekutieli false-discovery rate method.

Article Snippet: Viruses and in vivo analysis of infection Stocks of RSV strain A2 expressing green fluorescent protein (GFP) ( 21 ) or firefly luciferase (Luc) reporter genes were obtained from Viratree.

Techniques: Infection, Control, Standard Deviation, Immunohistochemical staining, Staining, Two Tailed Test

Journal: Acta Pharmaceutica Sinica. B

Article Title: Transient induction of actin cytoskeletal remodeling associated with dedifferentiation, proliferation, and redifferentiation stimulates cardiac regeneration

doi: 10.1016/j.apsb.2024.01.021

Figure Lengend Snippet: Transient transfection of the 2D2P genes using a non-integrating lentivirus system induces transient actin remodeling and a transient increase in cardiomyocyte proliferation in vitro . (A) Heatmap showing the expression patterns of actin polymerization genes ( Dmd , Ctnna3 , Palld , Ank2 , Fhod3 , and Diaph3 ) and actin depolymerization genes ( Cfl1 , Pfn1 , Tmsb10 , and Tmsb4x ) in CM0, CM2, CM3, CM6, and CM8 cells. (B) The ranking of maximum variation of expression (ΔExpression) in actin polymerization ( Dmd , Ctnna3 , Palld , Ank2 , Fhod3 , and Diaph3 ) and depolymerization ( Cfl1 , Pfn1 , Tmsb10 , and Tmsb4x ) genes. (C) Staining of DAPI (blue), pH3 (red), and α -actinin (green) in neonatal mouse ventricular myocytes (NMVMs) from postnatal Day 7 (P7) mice (C1). Scale bar, 20 μm. Quantification of pH3 + cardiomyocytes in NMVMs from P7 mice treated with siRNA for 48 h. ∗ P < 0.05 vs . Control. 6 repeats/∼600 cells in each group (C2). (D) Schematic diagram of the plasmid design for Tnnt2-2D2P polycistronic NIL. (E–H) Representative images of NMVMs from postnatal Day 7 mice treated with Control-NIL or Tnnt2-2D2P-NIL for 0, 4, or 8 days and immunostained for DAPI (blue), GFP (green), Ki67 (red), pH3 (red), EdU (red), and α -actinin (red/green) (E1–H1). Scale bar, 20 μm. Quantification of GFP + (E2), Ki67 + (F2), pH3 + (G2), and EdU + (H2) cardiomyocytes. ∗ P < 0.05 vs . Control-NIL. 5–6 repeats/400–600 cells in each group. (I, J) Representative images of NMVMs from postnatal Day 7 mice treated with Control-NIL or Tnnt2-2D2P-NIL for 0, 4, or 8 days and immunostained for DAPI (blue), F-actin (white), G-actin (green), and α -actinin (green/red) (I1–J1). Scale bar, 20 μm. Quantification of disorganized F-actin percentage (I2), and G-actin intensity (J2) in cardiomyocytes. ∗ P < 0.05 vs . Control-NIL. 100–120 cells in each group.

Article Snippet: To investigate the effects of Tmsb4x (or Tmsb4x-mutant ), Tmsb10 (or Tmsb10-mutant ), shRNA- Dmd , and shRNA- Ctnna3 on NMVMs, we used non-integrating lentiviruses encoding these genes with the Tnnt2 promoters and green fluorescent protein (GFP) reporter, which were obtained from Genechem (Shanghai, China).

Techniques: Transfection, In Vitro, Expressing, Staining, Control, Plasmid Preparation

Journal: Journal of Biomedicine and Biotechnology

Article Title: Gene Transfer into the Lung by Nanoparticle Dextran-Spermine/Plasmid DNA Complexes

doi: 10.1155/2010/284840

Figure Lengend Snippet: Fluorescent microscopy of GFP expression in COS-7 cell lines transfected with different gene carriers. Fluorescent cells were visualized at 48 h post-transfection. (a) Untreated, (b) naked DNA, (c) PEI/phMGFP, (d) D-SPM/phMGFP at ratio 12, (e) D-SPM/phMGFP at ratio 14, and (f) D-SPM/phMGFP at ratio16. (Magnification x10).

Article Snippet: Plasmid phMGFP carrying green fluorescent protein (GFP) reporter gene was purchased from Promega (Madison, WI, USA).

Techniques: Microscopy, Expressing, Transfection

Journal: Journal of Biomedicine and Biotechnology

Article Title: Gene Transfer into the Lung by Nanoparticle Dextran-Spermine/Plasmid DNA Complexes

doi: 10.1155/2010/284840

Figure Lengend Snippet: Side scatter (SSC) versus GFP fluorescent dot plot of transfected COS-7 cells showing positions of GFP-negative (R1) and GFP-positive (R2) population with the percentage gated of GFP-positive cells using flow cytometry analysis by using different gene carriers. Gene expression was measured at 48 h post-transfection. (a) Untreated, (b) naked DNA, (c) PEI/phMGFP, (d) D-SPM/phMGFP at ratio 12, (e) D-SPM/phMGFP at ratio 14, and (f) D-SPM/phMGFP at ratio16.

Article Snippet: Plasmid phMGFP carrying green fluorescent protein (GFP) reporter gene was purchased from Promega (Madison, WI, USA).

Techniques: Transfection, Flow Cytometry, Expressing