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This level of variation is small relative to that observed among livers from different animals in our larger sample.
Cytospin samples were sorted on MAS coated glass slides (Matsunami glass, Japan), followed by centrifugation at 2000rpm, 4°C for 5 min. For in vivo assays, C57BL/6J mouse livers from different developmental stages were embedded in Tissue-Tek® OCT compound (Sakura Finetek, Japan).
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Thorough examination of several liver sections from different mouse livers revealed that the majority of the labeling for PDE3B was localized to smooth ER (Figure 5).
Three different livers obtained from different animals were used, and from each liver three samples were extracted for polarimetric measurements, so a total of nine samples were investigated.
Frozen C57Bl/6 mouse livers from 3 different young (7 month) and 3 different older (17 month) mice were a gift from Dr. Rafael de Cabo (NIA).
Each sample contained several individual tumor metastases from the same animal or several liver sections from different areas of the liver for each animal.
The micrometastatic cells that were extracted to obtain RNA samples were captured by LCM from livers from three different areas (zones 1, 2 and 3).
In order to compare ATP and protein content in liver slices from different experiments, both ATP and protein were normalized on mass of liver slices.
The KSF has been investigated by our group in visualising pancreatic cancer, and liver metastases from different primary tumour sites, and we have consistently demonstrated the applicability of this technique [17, 18].
TMA from Figure 4a comprised 37 different PDAC samples, 17 lymph nodes and 10 liver metastases from different primary PDAC tumours metastases.
Interestingly a similar correlation was found when the liver transcriptome from different studies was compared, which in this case had a correlation of 53%.
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