Leukemia initiated by PMLRARα: the PML domain plays a critical role while retinoic acid–mediated transactivation is dispensable by Scott C. Kogan, Suk-hyun.

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Leukemia initiated by PMLRARα: the PML domain plays a critical role while retinoic acid–mediated transactivation is dispensable by Scott C. Kogan, Suk-hyun Hong, David B. Shultz, Martin L. Privalsky, and J. Michael Bishop Blood Volume 95(5):1541-1550 March 1, 2000 ©2000 by American Society of Hematology

Scott C. Kogan et al. Blood 2000;95:1541-1550 ©2000 by American Society of Hematology

Scott C. Kogan et al. Blood 2000;95:1541-1550 ©2000 by American Society of Hematology

Scott C. Kogan et al. Blood 2000;95:1541-1550 ©2000 by American Society of Hematology

Scott C. Kogan et al. Blood 2000;95:1541-1550 ©2000 by American Society of Hematology

Expression of the transgenes Expression of the transgenes.A, B, Whole cell lysates of bone marrow were subjected to denaturing PAGE and Western blotting using a rabbit polyclonal antihumanRARαF domain antibody. Expression of the transgenes.A, B, Whole cell lysates of bone marrow were subjected to denaturing PAGE and Western blotting using a rabbit polyclonal antihumanRARαF domain antibody. Signals corresponding to transgenically expressed PMLRARα and RARα proteins are indicated by arrows. Locations of size markers are indicated by lines. (A) Protein expression in nonleukemic bone marrow of the highest expressing MRP8-PMLRARα transgenic line (Tg556-PR) and in 2 of the MRP8-PMLRARαm4 mice, Tg4048-PRm4 (nonleukemic bone marrow) and Tg4099-PRm4 (leukemic bone marrow). 8% SDS-polyacrylamide. (B) Protein expression in nonleukemic bone marrow Tg556-PR and in the marrows of healthy MRP8-RARαm4 transgenic mice from 3 lines, Tg4142-Rm4, Tg4151-Rm4, and Tg4192-Rm4. 12% SDS-polyacrylamide. C, Immunofluorescence analysis of bone marrow neutrophilic cells, anti-RARαF antiserum and Hoechst 33258, 1300 × . Scott C. Kogan et al. Blood 2000;95:1541-1550 ©2000 by American Society of Hematology

Acute leukemia in MRP8-PMLRARm4 transgenic mice Acute leukemia in MRP8-PMLRARm4 transgenic mice.(A, C, E) Samples from control mice. Acute leukemia in MRP8-PMLRARm4 transgenic mice.(A, C, E) Samples from control mice. (B, D, F) Samples from leukemicPMLRARαm4 mice. (B) Founder 4099. (D) Transplanted leukemia 4048.2. (F) founder 4042. (A, B) Bone marrow, Wright's Giemsa stain, 500 × . (C, D) Bone marrow, Sudan Black B stain, 965 × . (E, F) Liver, Hematoxylin and eosin stain, 200 × . Scott C. Kogan et al. Blood 2000;95:1541-1550 ©2000 by American Society of Hematology

Variable surface marker expression in PMLRARm4leukemias Variable surface marker expression in PMLRARm4leukemias.Bone marrow cells were stained with Gr-1 and Mac-1 antibodies that recognize myeloid surface antigens. Variable surface marker expression in PMLRARm4leukemias.Bone marrow cells were stained with Gr-1 and Mac-1 antibodies that recognize myeloid surface antigens. Dead cells were eliminated from the analysis on the basis of staining with propidium iodide. (A) Control. (B) Leukemic mouse 4042 (C) Leukemic mouse 4099 (D) Transplanted leukemia 4048.2. Scott C. Kogan et al. Blood 2000;95:1541-1550 ©2000 by American Society of Hematology

Retinoic acid response of PMLRARm4 leukemias Retinoic acid response of PMLRARm4 leukemias.Leukemic mice were treated with placebo (A, C, E, G, I, K) or tRA (B, D, F, H, J, L). Retinoic acid response of PMLRARm4 leukemias.Leukemic mice were treated with placebo (A, C, E, G, I, K) or tRA (B, D, F, H, J, L). (A-D) PMLRARα expressing leukemia. (E-H)PMLRARαm4 leukemia 4099. (I-J) PMLRARαm4 leukemia 4042. (K-L) PMLRARαm4 leukemia 4048.2. (A, B, E, F, I-L) Bone marrow, Wright's Giemsa stain, 350 × . (C, D, G, H) Liver, Hematoxylin and eosin stain, 140 × . Effects of 11 days of tRA therapy are shown. Scott C. Kogan et al. Blood 2000;95:1541-1550 ©2000 by American Society of Hematology