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. 2017 Dec 28;8:96–105. doi: 10.1016/j.molmet.2017.12.013

Figure 1.

Figure 1

Characterization of the Lifeact-EGFP transgenic mouse islets. A) Schematic representation of the Lifeact-EGFP transgenic construct. Expression of the Lifeact peptide fused to EGFP is driven by CAG promoter [22]. Islets were isolated from Lifeact-EGFP adult mice and cultured either for 3D imaging or perifusion assays. B) Imaging of native Lifeact-EGFP (green) in live pancreatic islets. On the left, representative maximum intensity projection (MIP) of consecutive optical sections of a pancreatic mouse islet cultured for 12 h in the presence of glucose (16 mM); on the right, a single optical section (Single plane). Lifeact-EGFP signal labels all islet cells with a distribution more pronounced at the cellular edges (arrows) than along the cell faces (arrowheads). This is consistent with the labeling distribution observed in individual β-cells upon Lifeact-EGFP adenoviral infection of isolated mouse islets [29]. C) Representative single plane confocal image of whole-mount immunofluorescence staining of Lifeact-EGFP transgenic mouse islets for Phalloidin in red and Lifeact-EGFP in green. Insets show split channels (green and red) of boxed area; arrows indicate overlapping stainings. D) Representative single plane confocal image of whole-mount immunofluorescence of Lifeact-EGFP transgenic mouse islets for glucagon (Gluca, red) and insulin (Ins, blue), in green Lifeact-EGFP. Hoechst 33342 was used as nuclear counterstain. Insets show split channels of boxed area at higher magnification; stars indicate glucagon-positive cells which are negative for insulin. Bar, 10 um.