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. 2019 Mar 12;151(4):532–554. doi: 10.1085/jgp.201812294

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© 2019 Zawieja et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 4. — Recombination efficiency of SMMHC-CreER^T2, SMArTTa-tetOCre, smMHC^Cre/eGFP, and c-KitCreER^T2 lines with ROSA26mT/mG. (A) Representative immunofluorescence maximal projections of a fixed SMMHC-CreER^T2-ROSA26mT/mG IALV stained for GFP and SMA to determine recombination efficiency. (B) Representative immunofluorescence maximal projections of a fixed SMArTTa-tetOCre-ROSA26mT/mG IALV stained with antibodies for GFP and SMA to determine recombination efficiency. (C) Representative immunofluorescence maximal projections of a smMHC^Cre/eGFP IALV fixed and stained for GFP and SMA to determine recombination efficiency. (A–C) Comparisons of cell masks for SMA and GFP to highlight cells that were SMA-positive/GFP-negative and the merged maximal projection used to assess recombination efficiency. (D) Representative immunofluorescence maximal projections of a live and pressurized (2 cm H₂O) c-KitCreER^T2-ROSA26mT/mG IALV imaged for GFP (recombined cells) and td-tomato (not recombined). Images were taken at 20× objective magnification. (E) Representative immunofluorescent images of the intestine muscle layer of a c-KitCreER^T2-ROSA26mT/mG induced with our tamoxifen injection protocol and acquired using a fluorescence macroscope revealed recombination of the c-Kit⁺ interstitial cells of Cajal. RFP, red fluorescent protein (tdTomato); GFP, green fluorescent protein (EGFP).