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. 2022 Mar 18;13:1473. doi: 10.1038/s41467-022-29081-2

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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. 3 — a Bioluminescence imaging (BLI) analysis with ROI of mice bearing DFBM-727 (p16^INK4A positive) tumors at indicated weeks after treatment with control, tucatinib (PO 75 mg/kg, BID), abemaciclib (PO 75 mg/kg, QD), or Tuca + Abem. Left panel: representative bioluminescence images. Middle panel: quantification of the regions of interest (ROI) in each group mice at indicated imaging time points, Vehicle control (n = 5), Tucatinib (n = 4), Abemaciclib (wk0-4, n = 4; wk6, n = 2) or Tuca + Abem (n = 5). Mean ± SD. Right panel: Kaplan–Meier survivals of mice bearing DFBM-727. Dotted lines indicate treatment starting time. b BLI and survival analysis for DFBM-Ni8 (p16^INK4A IHC negative, CDKN2A CNV = 1) treated as in a. BLI right panel: Vehicle control (wk0,1 n = 5; wk2,3, n = 4; wk4, n = 3), Tucatinib (n = 5), Abemaciclib (wk0-3, n = 4; wk4, n = 3) or Tuca + Abem (wk0-2, n = 5; wk3,4, n = 4). Mean ± SEM. c BLI and survival analysis for DFBM-Ni17 (p16^INK4A IHC negative, CDKN2A CNV = 2) treated as in a. BLI right panel: Vehicle control (n = 3), Tucatinib (n = 3), Abemaciclib (wk0, n = 3; wk1,2, n = 2), or Tuca + Abem (n = 3). Mean ± SEM. d Multiplex immunofluorescence shows increase in p16^INK4A (control, n = 8 sections; Dox, n = 7 sections) and reduction in p-RB (control, n = 7 sections; Dox, n = 8 sections) after 3 days of doxycycline administration to DFBM-Ni17/tetp16^INK4A. Mean ± SD, Unpaired t-test. e BLI analysis with ROI of mice bearing DFBM-Ni17/tetp16^INK4A tumors at the indicated time with the treatment as indicated. Tucatinib (PO 75 mg/kg, BID), abemaciclib (PO 75 mg/kg, QD), doxycycline-diet (2500 ppm, ScottPharma). Control (n = 4), Dox (n = 4), Dox + Abem (n = 4), Dox + Tuc (wk0-3, n = 5; wk5, n = 4), Dox + Tuc + Abem (wk0-1, n = 5; wk2-5, n = 4). Mean ± SD. Source data are provided as a Source Data file.