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. 2018 Nov;15(4):389–399. doi: 10.20892/j.issn.2095-3941.2018.0122

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Copyright 2017 Cancer Biology & Medicine

This work is licensed under a Creative Commons Attribution-NonCommercial-Share Alike 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/

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PKM-knockout in 4T1 cells. (A) Schematic for PKM gene. Exon 1 is not shown here. # indicates sgRNA target position in PKM gene. (B) sgRNA target sequences for mouse PKM gene. sgPKM-#4 successfully knockout PKM gene. (C) Sequencing of PKM gene in 4T1/PKM-KO cells targeted by sgPKM-#4 as shown in (B). Two alleles are differentially damaged, one with 20bp deletion, the other containing 2bp loss. (D) Western blot of PKM2 in 4T1/Cas9, 4T1/PKM-KO and 4T1/KO+EGFP-PKM2 cells. 4T1/KO+EGFP-PKM2 cells are 4T1/PKM-KO cells expressing EGFP-PKM2. (E) (F) Relative lactate excretion and relative glucose uptake of 4T1/Cas9, 4T1/PKM-KO and 4T1/KO+EGFP-PKM2.