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. Author manuscript; available in PMC: 2021 Jan 13.
Published in final edited form as: Cancer Cell. 2020 Jan 13;37(1):71–84.e7. doi: 10.1016/j.ccell.2019.12.002

Figure 2. PDXK is required for leukemia disease maintenance in vivo.

Figure 2.

(A) Schematic showing the in vivo leukemia mouse model with sgRNA infected AML cells. (B-E) Nras(G12D)/MLL-AF9 cells infected with viruses encoding sgRNAs targeting Rosa26 (n=5) or Pdxk (Pdxk. sg1, n=7 and Pdxk.sg2, n=5) were transplanted to sub-lethally irradiated mice. On day 5 and day 14, luciferase signals were detected (B), quantified and represented as average and SD (C). ‘***’ represents p<0.001 of t-test between Rosa26 and Pdxk sgRNAs. Western blot was performed to measure the PDXK level of virus infected Nras(G12D)/MLL–AF9 cells before transplantation or taken from moribund mice, and the intensity was quantified and normalized to the corresponding Rosa26 sgRNA (D). “*” represents p<0.05 of t-test between Rosa26 and Pdxk sgRNAs (Rosa26 sgRNA, n=3; Pdxk.sg1 and 2, n=2). Survival curves are shown (E). “**” represents p<0.01 of Log-rank (Mantel-Cox) test between Rosa26 and Pdxk sgRNAs. (F) The schematic diagram shows the in vivo leukemia mouse model with shRNA infected AML cells. Doxycycline (Dox) was supplemented 5 days after transplantation. (G-J) Nras(G12D)/MLL–AF9 cells infected with viruses encoding shRNAs targeting Renilla luciferase (Ren, n=12) or Pdxk (Pdxk.sh1, n=13 and Pdxk.sh2, n=10) were transplanted to sub-lethally irradiated mice. On day 5 and day 14, luciferase signals were detected (G), quantified and represented as average and SD (H). ‘***’ represents p<0.001 of t-test between Ren and Pdxk shRNAs and ‘###’ represents p<0.001 of t-test between Pdxk shRNAs. Western blot was performed to measure the PDXK level of viruses infected Nras(G12D)/MLL–AF9 cells before transplantation or taken from moribund mice, and the intensity was quantified and normalized to the corresponding Ren shRNA (I). “**” represents p<0.01 of t-test between Ren and Pdxk shRNAs and “###” represents p<0.001 of t-test between Pdxk shRNAs (n=3). Survival curves are shown (J). ‘***’ represents p<0.001 of Log-rank (Mantel-Cox) test between Ren and Pdxk shRNAs and ‘##’ represents p<0.01 of Log-rank (Mantel-Cox) test between Pdxk shRNAs.