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. Author manuscript; available in PMC: 2019 Mar 11.
Published in final edited form as: Leukemia. 2018 Sep 11;33(3):671–685. doi: 10.1038/s41375-018-0248-0

Figure 7. Downregulating Notch signaling targets mitochondrial metabolism in Kras myeloid progenitor and precursor cells.

Figure 7

Lethally irradiated mice (CD45.1+) were transplanted with 2 ×106 splenocytes (CD45.2+) from KrasLSL G12D/+;Mx1-Cre (Kras) or KrasLSL G12D/+;Rosa26LSL DNMAML-GFP/+;Mx1-Cre (Kras; D/+) mice along with 2.5×105 competitor cells (CD45.1+). The control group was transplanted with with 1×106 bone marrow cells (CD45.2+) along with 2.5×105 competitor cells (CD45.1+). Three weeks after transplantation, Cre expression was induced using pI-pC injections as described in Methods. Recipients transplanted with control, Kras or Kras; D/+ cells were sacrificed 5 weeks after pI-pC injections. Donor-derived Lin− bone marrow cells were sorted using flow cytometry. Donor-derived cells are defined as CD45.2+ cells in control and Kras recipients or CD45.2+ GFP+ cells in Kras; D/+ recipients. (A) Oxygen consumption rates (OCR) were measured in the presence of the mitochondrial inhibitor (oligomycin, 1μM), the uncoupling agent (FCCP, 2.5 μM), and the respiratory chain inhibitor (rotenone, 1 μM). (B) Quantification of ATP-linked OCR, which is the calculated difference between the basal OCR level and the OCR level after oligomycin treatment. (C) Total cellular ATP concentrations were measured using the CellTiter Glo assay. (D) Quantification of extracellular acidification rates (ECAR). Data are presented as mean ± SD. * P<0.05, ** P<0.01; *** P<0.001.