Skip to main content
. 2018 Jul 30;16:65–75. doi: 10.1016/j.molmet.2018.07.011

Figure 1.

Figure 1

Insulin-like-5 rtTA mice faithfully report insulin-like-5 producing L-cells. (A) Scaled diagram of the BAC-transgene used to make Insl5-rtTA mice; Insl5-coding sequence was replaced with the reversed tetracycline activator (rtTA) coding sequence. (B) Cartoon illustrating doxycycline dependent labelling of Insl5-producing cells with either GFP or GCaMP6f. (C&D) Immunofluorescence based assessment of GFP induction in tissue sections (n = 3 mice) following in vivo doxycycline induction from Insl5-rtTA/TET-GFP mice. (E&F) Immunofluorescence based assessment of GCaMP6F induction in primary cultures (n = 3 mice) generated from Insl5-rtTA/TET-GCaMP6FΔCMV mice treated ex vivo overnight with doxycycline. Bars represent percentage of total cells stained within each of the following subgroups: INSL5 only (red), INSL5 and GFP co-stain (yellow) and GFP only (grey). (G) Representative FACS scatterplot illustrating the subpopulations of cells selected for downstream RT-qPCR. (H) Relative expression of Insl5, Gcg, and Pyy in FACS GFP+ve and GFP−ve cells following in vivo doxycycline induction (n = 6 mice). Bars represent mean 2ΔCT + SEM compared to β-actin. Statistical significance was assessed through ratio paired t-tests applied to 2ΔCT. ***p < 0.001.