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The initial water flow per unit area of microvessel wall [(Jv/S 0, where Jv is water flux and S is unit area of the microvessel wall] was calculated from the velocity of the marker cell after the vessel was occluded, the vessel radius, and the length between the marker cell position and the occlusion site (Fig. 1).
This was confirmed by flow cytometric analysis which showed the presence of double positive Gr-1 (myeloid differentiation antigen) and CD11b (macrophage and granulocyte marker) cell populations (71%) which compared closely to those derived from wt-iPSCs (85%) (Figure 3C).
For each marker, cell numbers were statistically compared between sham and treated animals using analysis of variance (ANOVA) followed by t-tests.
Assays were conducted using the Lister 427 single marker cell line or a TbUAP conditional null mutant grown in HMI9-T.
To confirm the cell surface localisation of LAMP1 on hypertrophic chondrocytes, cryosections from 6 day old mouse epiphyses were stained for LAMP1 together with the membrane marker, cell mask orange.
Ectoptic expression of mir-200c was carried out using an expression plasmid containing the mir-200c precursor expressed from an EF-1 α promoter and a puromycin resistance marker (Cell Biolabs).
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To select for cells that had recycled the amdSYM marker, cells were plated on SM containing 2.3 g L−1 fluoroacetamide (SM-FAc).
In agreement with others [24], this procedure yields 98% A2B5-positive (OPC marker), and 2% MBP-positive (mature OL marker) cells.
GFAP-positive (astrocyte marker) or Ox2A-positive (microglia marker) cells cannot be detected in cultures prepared in this manner.
Control mixes used 6.5 hr csaAMS2 cells and 6.5 hr nuclear marker cells.
Mice were treated with dox for one week, lungs were homogenized and CD45+ (leukocyte marker), F4/80+ (macrophage marker) cells were selected for further cell surface marker analysis.
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