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The first experiments were performed by using mouse cells.
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Specifically, by using mouse embryocarcinoma cells, we found that a dominant negative RARα mutant creates a concerted repression of both RARβ2 and its direct target CYP26A1, encoding the cytochrome P450 RA-specific hydrolase, which acts as a neuronal differentiation switch in these cells [8], [10].
In this manuscript, the authors analyzed the role of Oct6 in during neuronal commitment and differentiation by using mouse ES cell differentiation system.
The results from human studies were finally confirmed in vitro with human muscle cell experiments and by using mouse mature muscle cells in culture.
They accomplished this by using mouse embryonic stem cells to manipulate the mouse genome at the very start of development.
Tregs were analyzed by using mouse regulatory T cell Staining Kit (eBioscience).
Bioactivity of vIL-10 in the fusion protein was tested with a cell-proliferation assay by using mouse MC-9 mast cells as described [ 18].
To investigate the significance of protein changes under disease conditions, we have chosen a more systematic and simplified approach by using mouse embryonic stem (ES) cells with highly defined modifications in a controlled environment.
For in vitro T-cell assays, spleen cells from naive mice were depleted from CD3+ cells by using mouse anti-CD3ε antibodies (clone 17A2; BD Biosciences) and sheep anti-rat IgG Dynabeads at a ratio of two beads per cell (Invitrogen) in combination with Dynal MPC-1 Magnetic Particle Concentrator (Dynal A.S). and were used as APCs with a purity of more than 95% CD3− cells.
The electrospun nanofibrous membrane was evaluated in vitro by using mouse fibroblasts (L929) as reference cell lines.
In vitro cytotoxicity was investigated by using mouse embryonic fibroblast (BALB/3T3) and N2a cells for any embryo- and neuro-toxic effects; respectively.
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