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Cultures were then treated with lapatinib, an IKKβ inhibitor,38 or an inhibitor of the CCL2 receptor, CCR2 (RS504393), and macrophages isolated from mammary glands of MMTV-HER2 mice were added to the cultures.
For bronchoalveolar lavage assays, an additional 5 mice were added for each group.
Purified Gr-1+CD11b+ cells from IL-6-treated mice were added to CFSE-labeled T cells stimulated with anti-CD3.
Splenocytes from individual mice were added in duplicates and incubated for 48 h at 37°C, 5% CO2 with 5 µg of OVA.
Splenocytes isolated from these mice were added in serial dilution to DNA-coated plates and incubated for 12 hours at 37°C.
In some wells, FACS-sorted CD4+6.5+CD25− or CD25+ T cells obtained from dtg mice were added to CD4+6.5+CD25− from stg mice at a ratio of 1∶1.
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A standard serum composed of a mixture of sera from the arthritic mice was added to each plate in serial dilutions and a standard curve was constructed.
A standard serum composed of a mixture of sera from arthritic mice was added to each plate in serial dilutions and a standard curve was constructed.
In animal studies impaired capacity to reduce surface tension occurred when BAL fluid from RSV-infected BALB/c mice was added to calf lung surfactant extract [ 25].
This medium was made sterilized by pressure passage through 0.22 μm membrane filters (Millipore, Germany) and then 10 mL (10%) urine (filtered and autoclaved of both sheep and mice) was added.
When purified CD8+ T cells of NP- or P-mice were added to their respective CD4+ T cells a significant inhibition of Ag85B-mediated IFN-γ secretion was observed (Fig. 1 A and B), indicating that CD8+ T cells exert suppressing activity on antigen-specific CD4+ T cells.
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