D14 D 10 D 7. Untreated CYTOXAN 5-FU
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1 Dipeptidylpeptidse Negtively Regultes Colony Stimulting Fctor Activity nd Stress Hemtopoiesis. Hl E. Broxmeyer, Jonthn Hoggtt, Hether A. O Lery, Chrlie Mntel, Brhmnnd R. Chitteti, Scott Cooper, Steven Messin-Grhm, Gio Hngoc, Sherif Frg, Sr L. Rohrbugh, Xun Ou, Jennifer Speth, Louis M. Pelus, Edwrd F. Srour, nd Timothy B. Cmpbell cgy cgy Colonies per femur (thousnds) Untreted CFU-GEMM D D D Untreted D D D Untreted D D D CBL/ (WildType) Dpp -/- b Colonies per femur DAY CFU-GEMM Untreted cgy FU Untreted cgy FU Untreted cgy FU CBL/ (WildType) Dpp -/- c d Colonies per femur (thousnds) C: Untreted Dpp -/- : Untreted C: CTX-D Dpp -/- : CTX-D C: CTX-D Dpp -/- : CTX-D C: CTX-D Dpp -/- : CTX-D C: CTX-D Dpp -/- : CTX-D C: Untreted Dpp -/- : Untreted C: CTX-D Dpp -/- : CTX-D C: CTX-D Dpp -/- : CTX-D C: CTX-D Dpp -/- : CTX-D C: CTX-D Dpp -/- : CTX-D CYTOXAN -FU Percent progenitors in S-phse Untreted D D9 D D D D D Untreted D D9 D D D D D CFU-GEMM Untreted D D9 D D D D D CBL/ (WildType) Dpp -/- Supplementl Figure. In vivo role of DPP in recovery of hemtopoiesis fter stress. ) Absolute numbers of HPC in WT vs. Dpp-/- mice prior to nd fter or cgy g-irrdition. b) Absolute numbers of HPC in WT vs. Dpp-/- mice prior to nd dys fter cgy g-irrdition or mg/kgbwt -FU i.v. c) In vivo role of Dpp in recovery of bsolute numbers of nd fter tretment of CBl/ control nd Dpp-/- mice with cyclophosphmide (Cytoxn; mg/kgbwt i.p.)., significnt (p<.) compred to CBl/ control of tht dy., not done. d) Cycling sttus (% HPC in S-Phse) prior to nd fter tretment of WT or Dpp-/- mice with mg per Kgbwt -FU i.v. nd mg per Kgbwt CytoxnR i.p. Results shown re the men ±s.e.m. for mice per group, ech mouse being ssessed individully., significnt (p<.) chnge compred to WT vlue of tht dy. Nture Medicine doi:./nm.99
2 Cells per femur (thousnds).x.x.x.x.x (CD-LSK).x.x.x.x.9x.x D D D D D.x.x.x.9x.x.9x.x (CD+LSK).x.9x MPP.x.x.x.x.x.x.x.x D D D D D.x.x D D D D D.x.9x.x MEP.x.x.9x.x.x D D D D D Recovery fter cgy x.x (CD+LSK).x.x.x.x.x.x.x.9x D D D D D.x.x.x.x.x.9x.x.x (CD-LSK).x.x CLP.x.x.x.x D D D D D D D D D D.x.x GMP.x D D D D D b Cells per femur (thousnds).x.x.x.x x (CD-LSK).x.x.x x.x D D D D D.x.x.x.x (CD+LSK).x.9x.x.x.x D D D D D.x.x.x MPP.x.x.x.x.x D D D D D.x.9x.x.x MEP.x.x.x.x.x D D D D D Recovery fter FU x.x.x.x (CD+LSK).x.x.x.x.x D D D D D.x.x (CD-LSK).x.x.x.x.x.x.9x D D D D D.x.x.x.x CLP.x.x.x.x.x D D D D D.x.x x x GMP x D D D D D c Cells per femur (millions) Myeloblst D D D D D Met + Bnds D D D D D Recovery fter cgy Promyelocytes nd Myelocytes PMN D D D D D D D D D D Myeloblst WT WT+Sitgliptin Dpp -/- d Cells per femur (millions) D D D D D Met + Bnds D D D D D Recovery fter FU Promyelocytes nd Myelocytes D D D D D PMN D D D D D Supplementl Figure. Recovery of phenotypiclly defined HSCs nd HPCs ( nd b) nd morphologiclly recognizble cells of the myeloid linege (c nd d) fter stress. Recovery from cgy rdition ( nd b) nd FU (b nd d). X = fold chnge from indicted mice. Sttistics re compred to WT of tht dy. p<.. = not significnt (p>.) Nture Medicine doi:./nm.99
3 . Tretment: Dy-: Sitgliptin Dy-: Sitgliptin; hrs lter: FU or cgy b. Recovery from FU shown s dys fter tretment i WBC ii Neutrophils iii Lymphocytes Percent bseline pre-stress iv v vi Percent bseline pre-stress Percent bseline pre-stress Percent bseline pre-stress Percent bseline pre-stress Percent bseline pre-stress Pltelets WT(control) WT (Sitgliptin) Dpp -/- c. Recovery from cgy -irrdition shown s dys fter tretment i WBC ii Neutrophils iii Lymphocytes iv v vi Pltelets Supplementl Figure. Recovery of peripherl blood cells. ) Tretment of mice. b) Recovery from FU. c) Recovery from cgy. Results re bsed on - mice/group for WT nd mice/group for WT plus sitgliptin, or Dpp-/- mice with results expressed for ech mouse group s percent of tht mouse stedy-stte pre-fu or -cgy cell number. Sttistics for enhnced recovery t ech dy post-stress compred Dpp-/- to WT (, p<.) or WT plus sitgliptin to WT (, p<.). WBC, white blood cells;, red blood cells. Nture Medicine doi:./nm.99
4 . Model: T= FU T=-hr Sitgliptin T=hr Sitgliptin b. Recovery from FU shown s dys fter tretment i ii iii WBC Neutrophils Lymphocytes iv v vi Pltelets WT(control) WT (Sitgliptin) Dpp -/- Supplementl Figure. Recovery of peripherl blood cells in mice treted with FU. ) Tretment of mice with sitglipitin, which vries from the tretment shown in S. Fig. b) Cell recovery fter FU. Cohorts of mice per group for WT nd WT plus sitgliptin, nd mice per group for Dpp-/- were not bled more often thn every dys. It is not cler why there ws such lrge increse in cell counts for WT (control) mice round - dys fter FU in S. Fig. b compred to tht seen in experiments shown in S. Fig.. Nture Medicine doi:./nm.99
5 Amino cid sequences for bsence or presence of puttive DPP clevge sites on murine nd humn cytokines Growth Fctor GM-CSF G-CSF IL- EPO M-CSF SCF FL Species Full length N-terminus Predicted Predicted Truncted N- Terminus Mss Mss Humn APARSPSPSTQPWEH,9 ARSPSPSTQPWEH, Mouse APTRSPITVTRPWKH, TRSPITVTRPWKH,9 Humn TPLGPASSLPQSFLL, LGPASSLPQSFLL, Mouse VPLVTVSALPPSLPL,9 LVTVSALPPSLPL, Humn APMTQTTSLKTSWVN, MTQTTSLKTSWVN,9 Mouse ASISGRDTHRLTRTL, N/A N/A Humn APPRLICDSRVLERY ~ kd PRLICDSRVLERY ~ kd Mouse APPRLICDSRVLERY ~ kd PRLICDSRVLERY ~ kd Humn EEVSEYCSHMIGSGH, N/A N/A Mouse KEVSEHCSHMIGNGH,9 N/A N/A Humn EGICRNRVTNNVKDV, N/A N/A Mouse KEICGNPVTDNVKDI,9 N/A N/A Humn TQDCSFQHSPISSDF, N/A N/A Mouse GTPDCYFSHSPISSN, N/A N/A This my be in dimer form Glycosyltedprotein Supplementry Tble Nture Medicine doi:./nm.99
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