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This variable uptake of PSMA-targeted radioligands in mouse spleen was already observed by us and others [13, 30 32].
7 to 10 days after the second DNA injection, mouse spleen was smashed with a syringe plunger in a 70 µm cell strainer (100 µm Nylon, BD).
The number of mononuclear cells per mouse spleen was counted on hemocytometer and the absolute number of a cell subset was calculated based on the percentage of cells stained for the appropriate markers [19].
6 days after transfer, enriched CD11c cells from the whole mouse spleen was analyzed by flow cytometry with 0.7∼3.0 × 10 events collected.
Consistent with the increased numbers of MZ B cells observed in flow cytometry, the MZ area of Gpr97 −/− mouse spleen was larger than that of WT controls.
For harvesting splenocytes, mouse spleen was aseptically dissected after euthanization and was homogenized between the frosted ends of the glass slides in the DMEM.
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As shown in Fig. 5A, Tregs in mouse spleen were effectively depleted by CD25 antibody.
Although PSMA (GCPII -expression in mouse spleen is documented on the mRNA-level [23], it is not detectable on the protein level [22].
The massive appearance of IMERV in the FDC network of the mouse spleen is a remarkable finding by itself.
EDCs did not express the pan-leukocyte marker CD45 or the macrophage marker MAC-1 (Figure 7e h; sections of mouse spleen were used as the positive control).
Therefore, to determine the possibility of glycosylation, PLD4 proteins from mouse spleen were analyzed by enzymatic deglycosylation (because of difficulty with analyzing trace amounts of endogenous PLD4 protein expressed in the cerebellum).
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