Journal: BMC Genomics

Article Title: The global gene expression outline of the bovine blastocyst: reflector of environmental conditions and predictor of developmental capacity

doi: 10.1186/s12864-021-07693-0

Figure Lengend Snippet: Molecular signature associated with developmental capacity of in vitro derived embryos. The heatmap indicates the expression patterns of the top 58 differentially expressed genes between CVT and NVT. Numbers 1, 2, 3 indicate three biological replicates hybridization whereas 1 μg of labelled Cy-5 labelled CVT samples were hybridized with 1 μg of Cy-3 labelled NVT samples. Numbers 4, 5, 6 indicate dye-swaps in which 1 μg of labelled Cy-3 labelled CVT samples were hybridized with 1 μg of Cy-5 labelled NVT samples

Article Snippet: Two microgram of amplified RNA from each sample (CVO, CVT, NVO and NVT) was mixed with either 1 μl of Cy-3 or Cy-5 ULS fluorescent labelling kit (Kreatech Diagnostics, Amsterdam, Netherlands) and incubated at 85 °C for 30 min. Unincorporated Cy-3 and Cy − 5 dyes were removed using the PicoPure RNA extraction kit (Applied Biosystems).

Techniques: In Vitro, Derivative Assay, Expressing, Hybridization

Journal: Journal of biosciences

Article Title: Circulating nucleic acids damage DNA of healthy cells by integrating into their genomes

doi:

Figure Lengend Snippet: Cellular entry, nuclear uptake and chromosomal association of fluorescently labelled DNAfs and Cfs. NIH3T3 cells (10×104) were treated with DNAfs labelled with ULS (red) and Cfs dual-labelled with ULS (red) and ATTO-TEC (green) (10 ng DNA in all experiments). (A) Intracellular fate of DNAfs at 30 min as analysed by LSCM. Numerous fine fluorescent particles are seen in the cytoplasm and in the nucleus. DIC, DAPI and ULS pictures are represented in different panels. (B) Intracellular fate of Cfs at 6 h as analysed by LSCM. Presence of dual-labelled Cfs in the cytoplasm and nuclei are clearly seen. The red and green signals appear yellow in colour when the images are overlapped. (C) Kinetics of nuclear uptake of fluorescently labelled DNAfs and Cfs analysed by LSCM. Fifty nuclei were analysed at each time-point and the percentage of positive nuclei was recorded. Nuclei containing at least two fluorescent spots were considered as positive. (D and E) Association of fluorescently labelled DNAfs (D) and Cfs (E) with chromosomes of treated cells. NIH3T3 cells were treated with labelled DNAfs and Cfs and metaphase spreads were prepared 6 h after treatment and analysed by fluorescence microscopy. Note that the labelled DNA particles are considerably smaller in size than Cfs particles.

Article Snippet: RNA was quantified using Nanodrop 1000 spectrophotometer (Thermo Scientific, USA) and labelled using ULS™ microRNA Labeling Kit (With Cy3) (Kreatech Diagnostics, The Netherlands.

Techniques: Fluorescence, Microscopy