Journal: Frontiers in Microbiomes

Article Title: A multi-strain human skin microbiome model provides a testbed for disease modeling

doi: 10.3389/frmbi.2025.1473292

Figure Lengend Snippet: (A) Relative abundance of the six applied consortia species in applied inoculum on Day 0 as measured by CFU. (B) Relative Abundance of the six strains harvested from skin tissue on Day 4 and Day 7 as measured by qPCR of microbiome genomic DNA. Absolute abundance values are reported in Table 4 . Each bar represents a single Transwell tissue within an experiment. (C) Alpha diversity was interpreted using Shannon Diversity. Significance was determined by unpaired t-test with Welch correction. Ns, not significant.

Article Snippet: To produce single strain gDNA template for qPCR standard curve generation, a modified protocol of New England Biolab’s Genomic DNA Purification Kit (NEB #T3010) was utilized on each strain used in the study.

Techniques:

Journal: Frontiers in Microbiomes

Article Title: A multi-strain human skin microbiome model provides a testbed for disease modeling

doi: 10.3389/frmbi.2025.1473292

Figure Lengend Snippet: (A) Relative abundance of applied inoculum on Day 0 as measured by CFU. (B) Relative abundance of the six strains on Day 4 and Day 7 as measured by strain specific qPCR assays of microbiome genomic DNA extracted from tissues with (AD+) or without (AD-) the addition of an AD cytokine mix. (C) Alpha Diversity of relative abundance as interpreted by a Shannon Diversity calculation. (D) Absolute abundance of strains harvested from tissue as measured by qPCR. Each bar represents microbiome harvested from single Transwell tissue within an experiment. Numerical values of absolute abundance are reported in Table 5 . Significance was determined by Multiple unpaired t-test with Welch correction, *p<.05. Ns, not significant.

Article Snippet: To produce single strain gDNA template for qPCR standard curve generation, a modified protocol of New England Biolab’s Genomic DNA Purification Kit (NEB #T3010) was utilized on each strain used in the study.

Techniques:

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