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. 2022 Feb 28;13:1090. doi: 10.1038/s41467-022-28619-8

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Schematic of the XTR cassette inserted within the first intron of Lkb1 in the eXpressed, Trapped, and Restored conformations. The XTR cassette is composed of an inverted gene trap consisting of an Ad40 splice acceptor upstream of eGFP. The gene trap is flanked by heterotypic loxP sites to allow for stable Cre-mediated inversion, which results in the truncation of wild-type Lkb1 transcripts. The gene trap is nested between two FRT sites to enable FLPo-ER^T2 -mediated deletion in the presence of tamoxifen, which results in the restoration of wild-type Lkb1 transcripts and LKB1 protein. b Assessment of the impact of Lkb1 restoration on tumor burden. Lung tumors were initiated in KT, KT;Lkb1^XTR/XTR, and KT;Lkb1^XTR/XTR;FLPo-ER^T2 mice. At 6 weeks post-initiation, tumor-bearing mice were treated for 6 weeks with corn oil vehicle or tamoxifen prior to analysis. IFU infectious units, VEH vehicle, TAM tamoxifen. c Representative fluorescence (top) and hematoxylin–eosin (H&E) staining (bottom) images of tumor-bearing lungs from KT, KT;Lkb1^XTR/XTR, and KT;Lkb1^XTR/XTR;FLPo-ER^T2 mice treated with vehicle or tamoxifen. Lung lobes within fluorescent images are outlined in white. Top scale bars = 5 mm. Bottom scale bars = 2 mm. d, e Tumor area (d) and tumor size (e) as assessed by histology for tumor-bearing KT, KT;Lkb1^XTR/XTR, and KT;Lkb1^XTR/XTR;FLPo-ER^T2 mice at 12 weeks after tumor initiation, following 6 weeks of treatment with vehicle or tamoxifen. In d, each dot represents a mouse, while each dot in e corresponds to a tumor. Red crossbars indicate the mean. In d, KT-vehicle, n = 3 mice; KT-tamoxifen, n = 3 mice; KT;Lkb1^XTR/XTR-vehicle, n = 5 mice; KT;Lkb1^XTR/XTR-tamoxifen, n = 5 mice; KT;Lkb1^XTR/XTR;FLPo-ER^T2-vehicle, n = 4 mice; KT;Lkb1^XTR/XTR;FLPo-ER^T2-tamoxifen, n = 4 mice. In e, KT-vehicle, n = 11 tumors; KT-tamoxifen, n = 14 tumors; KT;Lkb1^XTR/XTR-vehicle, n = 147 tumors; KT;Lkb1^XTR/XTR-tamoxifen, n = 135 tumors; KT;Lkb1^XTR/XTR;FLPo-ER^T2-vehicle, n = 167 tumors; KT;Lkb1^XTR/XTR;FLPo-ER^T2-tamoxifen, n = 19 tumors. One tissue section per mouse was analyzed. P values calculated by two-sided unpaired t test. VEH vehicle, TAM tamoxifen.