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Stimulation of murine islets with 11.6 nM MCP-1 or 16 mM glucose for 24 h, all significantly increased the levels of proamylin (∼8 kDa) and the intermediate form (∼6 kDa) of amylin (Figure 2A and 2B).
Incubation of murine islets with 16.6 mmol/l glucose resulted in a 9 10-fold increase in insulin secretion and the magnitude of this response was not influenced by prior incubation with 100 ng/ml Ptx (Fig. 7a).
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These results demonstrate scaffolds support murine islet transplantation with high efficiency, and feasibility studies in large animals support continued pre-clinical studies with scaffolds as a platform to control the transplant microenvironment.
Surprisingly, CCR2 mRNA was not detectable in both MIN6 cells and murine islets, and treatment of MIN6 cells with 11.6 nM MCP-1 for up to 9 hours had no effect on CCR2 expression (Figure 3A).
After precipitation of chromatin from SJ β-cells and murine islets by anti-Insm1 antibodies, we observed substantial re-precipitation with anti-Neurod1 or anti-Foxa2 antibodies (Fig 7A D).
Studies of optimal culture conditions demonstrated that RPMI 1640 with serum maintains or augments glucose-stimulated insulin secretion in murine islets [ 30].
Similar to human islets, murine islets stain positive for tetranectin.
Our studies showed that MCP-1 increased proamylin and the intermediate form of amylin in murine islets.
Murine islets were isolated according to the method described by Ricordi et al. in 1988 with slight modifications as described earlier [ 14, 15].
Murine islets were isolated using a modified version of a previously published islet isolation protocol (Rostambeigi et al., 2011).
Murine islets are typically composed of a β-cell core and a non-β-cell mantle.
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