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. 2019 Jul 22;38:322. doi: 10.1186/s13046-019-1322-9

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© The Author(s). 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 3 — Identification and validation of oncogenic signaling pathways that confer resistance to Palbociclib treatment. a Reactome pathway analysis with sgRNA candidates that confer resistance to Palbociclib. b,c Transcriptional activation of the 5 candidate genes was confirmed by qPCR in T24 SAM cells transduced with the according sgRNAs. d Detection of cell growth using a SRB cell proliferation assay after 4 days of Palbociclib (1000 Nm) treatment. Bar graph indicates the absorbance relative to NTC (*, P < 0.05; unpaired t-test), (e) 10 day clonogenic assay under Palbociclib (1000 nM) treatment. (Surviving fraction = colony number/seeded cell number in %; * means P < 0.05; unpaired t-test). Data represent the mean ± SD of 3 replicates