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This estimate revealed that when BAPN treatment was initiated either on d −1 or d 0, the average number of metastatic sites per mouse decreased by 44% (P = 0.0028), and 27% (P = 0.0284), respectively, as compared to the 'no BAPN' controls (Fig. 3C).
(C) Number of ZsGreen-positive (ZsG+) metastatic sites per mouse intracardiac (IC -injected wIC -injected MDA-MB-231 cells in C, E, or E+P states.
Cohorts of mice were given injections in the anterior flank (2 sites per mouse) with 10 tumor cells suspended in 0.1 ml of serum free medium.
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With one high affinity binding site per mouse rDNA, and with CTCF preferring non-methylated (and thus active) rDNA repeats, it is expected that only a small number of DNA-bound nucleolar CTCF molecules would be present.
Subconfluent cells in exponential growth were harvested with trypsin, washed twice with PBS and resuspended in PBS at 5 × 10 cells ml−1 and 0.1 ml of cell suspension was injected subcutaneously into the right flank, one site per mouse.
For subcutaneous xenograft assays with shRNA-transduced cells, nude mice (Taconic) were injected orthotopically into the mammary fat pad with 1.0 × 10 SUM190 or SUM185 cells, or subcutaneously with 1.0 × 10 BT-474 cells in serum-free medium with 50% Matrigel (BD Biosciences), one injection site per mouse.
Green fluorescent protein (GFP) tagged-MDA-MB-231 (ERα, ERβ or a metastatic variant of GFP-MDA-MB-435 (ER cells (~ 1 × 10) in Matrigel (BD Biosciences, San Jose, CA) were injected into the fourth right mammary fat pad (only one site per mouse) under isofluorane inhalation to produce orthotopic primary tumors as described in [ 26].
Three or four sites on pancreatic parenchyma per mouse were injected with 2 μL of pmaxGFP + TB and then electroporated.
In order to characterize which populations of cells were effectively targeted by B11-hCGβ, hMR-Tg and WT mice were injected s.c. in the paws with 5 µg of Alexa647-labeled B11-hCGβ in 50 µl saline per site (total 20 µg per mouse) in the presence or absence of adjuvant as indicated.
A 80- μl cell suspension containing 1 × 10 cells was subcutaneously injected into the hind legs of each mice, thus producing two sites of MCF-7 tumor per mouse.
A 80- μl cell suspension containing 1 × 10 cells was subcutaneously injected into the hind legs of each mice, thus producing a site of MCF-7 tumour per mouse.
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