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To substantiate the finding of higher insulin receptor phosphorylation in DEP-1 ASO-treated mice, liver sections were subjected to Proximity Ligation Assays (PLA) using co-incubation of antibodies directed against the insulin receptor and phosphotyrosine residues.
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(a) H&E staining of mice liver section on day 1 and 7 post PBS and hMSG-EpiPCs injection (White arrows: erythrocyte diapedesis, yellow arrows: diseased hepatocytes).
To identify this mechanism, we stained mouse liver sections to detect markers of various cell populations.
We validated these model findings with additional experiments on mouse liver sections.
All paraffin-embedded mouse liver sections were scanned with a digital slide scanner (Hamamatsu, Nanozoomer 2.0-RS) and files were analysed with the NDP viewer software.
For immunolocalization in liver tissues, paraffin-embedded mouse liver sections (5 µm) were dried for 1 h at 58 °C, followed by antigen retrieval and incubation with primary antibody (anti-cleaved caspase-3, Cell Signaling, #9661; anti-F4/80, eBioscience, #12 4801 80) in a Ventana automated instrument (Ventana Medical Systems, USA).
(C) Photomicrographs of representative H&E staining of mouse liver sections 6 h post-ConA injection (original magnification, 200× and 400×).
In order to determine its binding activity against native hepcidin we first performed immunohistochemistry on paraffin embedded mouse liver sections.
Four weeks following cell transplantation, the transplanted DiI-labeled hAMCs were detected and expressed human α-fetoprotein and albumin in mouse liver sections (Fig. 6).
Paraffin sections were processed similarly as mouse liver sections.
Formalin-fixed microtissues were paraffin embedded, sectioned, and processed similarly as mouse liver sections.
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