Journal: Nature Communications

Article Title: Muscle-specific gene editing therapy via mammalian fusogen-directed virus-like particles

doi: 10.1038/s41467-025-64200-9

Figure Lengend Snippet: a Schematic diagram illustrating the production of MuVLP Cas9 . b The levels of muscular fusogens, and Cas9 in VLPs, MuVLPs, VLP Cas9 , and MuVLP Cas9 . c ELISA quantification of Cas9 molecules per MuVLP Cas9 . The values and error bars represent the means ± s.d. of n = 3 technical replicates. d Scheme of MuVLP Cas9 or other controls cocultured with six different cell lines for detecting the specificity of MuVLP Cas9 for gene editing. e Immunofluorescence visualization of the Cas9 protein in cells cocultured with VLP Cas9 or MuVLP Cas9 . Red: anti-Cas9 antibody, green: F-actin labeled with Alexa Fluor 488 phalloidin, blue: nuclei labeled with DAPI. Scale bar, 20 μm. f Detection of exon 4 excision by genomic PCR in iC2C12 cells transfected with MuVLP Cas9 or other controls. The unedited genomic product is 883 bp long, and the gene-edited product (asterisk) is 445 bp long. g Sanger sequencing of amplicons confirmed the deletion of exon 4 and the generation of a fused intron 3/4 in genomic DNA from iC2C12 cells transfected with MuVLP Cas9 . The data are presented as the means ± s.d. of n = 3 biological replicates ( c ) or are representative of three independent experiments ( b , e – g ). Source data are provided as a Source Data file.

Article Snippet: Genomic DNA was extracted from tissues and cultured cells using a Genomic DNA Extraction Kit (D0063, Beyotime) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Immunofluorescence, Labeling, Transfection, Sequencing

Journal: ACS Nano

Article Title: Graphene Oxide Attenuates Th2-Type Immune Responses, but Augments Airway Remodeling and Hyperresponsiveness in a Murine Model of Asthma

doi: 10.1021/nn406454u

Figure Lengend Snippet: GO stimulates accumulation of chitinases (AMCase and CHI3L1) in the lungs of mice. Levels of chitinase 3-like 1 (CHI3L1) and acidic mammalian chitinase (AMCase) were evaluated in BAL fluid of mice 7 days after exposure to GO (A, B) or supernatants from cultured BAL macrophages: (A) AMCase; (B) CHI3L1. Alveolar macrophages were isolated from BAL fluid of mice exposed to GO or vehicle (24 h after exposure). The adherent cells (macrophages) were collected and seeded at a concentration of 2 × 10 5 cells/mL in 96-well plates in complete medium in the presence of recombinant mouse IL-4 (1000 U/mL). Forty-eight hours later, culture supernatants were collected and used for chitinase assay. * p < 0.05 vs PBS ( N = 2).

Article Snippet: The activity of AMCase was measured fluorimetrically on the basis of enzymatic hydrolysis of the 4-methylumbelliferyl-β- d - N,N ′, N ″-triacetylchitotriose (4-MU-chitotrioside, a substrate for endochitinase) with production of 4-methylumbelliferone (4-MU) using CycLex acidic mammalian chitinase fluorometric assay kit (MBL International, Woburn, MA, USA).

Techniques: Cell Culture, Isolation, Concentration Assay, Recombinant

Journal: ACS Nano

Article Title: Graphene Oxide Attenuates Th2-Type Immune Responses, but Augments Airway Remodeling and Hyperresponsiveness in a Murine Model of Asthma

doi: 10.1021/nn406454u

Figure Lengend Snippet: Molecular modeling of GO binding to AMCase and CHI3L1. The two predicted binding poses of GO, site1 and site2, in each case are represented as sticks and colored in gray and magenta, respectively: (A) AMCase; (B) CHI3L1. The structures of chitinases are represented as cartoons and colored red/blue to emphasize the N/C terminus. To highlight the active binding site cavity of chitinases, bound ligand/cavity space is represented as spheres. The oxygen and hydrogen atoms corresponding to the organic molecules—such as GO and small molecules bound to CHI3L1—are colored in red and white, accordingly.

Article Snippet: The activity of AMCase was measured fluorimetrically on the basis of enzymatic hydrolysis of the 4-methylumbelliferyl-β- d - N,N ′, N ″-triacetylchitotriose (4-MU-chitotrioside, a substrate for endochitinase) with production of 4-methylumbelliferone (4-MU) using CycLex acidic mammalian chitinase fluorometric assay kit (MBL International, Woburn, MA, USA).

Techniques: Binding Assay

Journal: ACS Nano

Article Title: Graphene Oxide Attenuates Th2-Type Immune Responses, but Augments Airway Remodeling and Hyperresponsiveness in a Murine Model of Asthma

doi: 10.1021/nn406454u

Figure Lengend Snippet: GO exposure inhibits the chitinolytic activity of AMCase. (A) Time course of chitinolytic activity in the BAL fluid of mice 24 h after the last OVA challenge ( i . e ., on day 31). (B) Dose-dependent changes in the enzymatic activity of recombinant human AMCase (hAMCase) upon addition of GO. (C) Decrease in the reaction rate of the enzyme upon addition of various concentrations of GO. A dose-dependent decrease in hAMCase upon addition of a known inhibitor, bisdionin C (IC 50 : 20 μM), is shown as an inset for comparison purposes. Data are shown as mean ± SEM, * p < 0.05 vs PBS treatment, and p < 0.05 vs OVA/ALL treatment ( N = 3).

Article Snippet: The activity of AMCase was measured fluorimetrically on the basis of enzymatic hydrolysis of the 4-methylumbelliferyl-β- d - N,N ′, N ″-triacetylchitotriose (4-MU-chitotrioside, a substrate for endochitinase) with production of 4-methylumbelliferone (4-MU) using CycLex acidic mammalian chitinase fluorometric assay kit (MBL International, Woburn, MA, USA).

Techniques: Activity Assay, Recombinant