Fig. 4.
Fig. 4. BSAP/Pax5A-expressing EML cells fail to expand to myeloid growth factors. (A) EML cells were induced to differentiate into myeloid cells as described in Fig 2A and then replated in 1% methylcellulose with 250 U/mL muGM-CSF at 1.5 × 105 cells per 3.5-cm plate. On day 9, the colonies were aspirated and analyzed by flow cytometry to determine the mean number of CD11b+/Gr-1+ cells per colony. The average among 3 control EML/Hyg clones and the average among 3 BSAP/Pax5A-expressing EML clones are shown and were found to be significantly different from each other using the heteroscededastic 1-tailed t-test (P < .001). (B) EML cells induced to differentiate as described in Fig 2A were replated in suspension culture in the presence of 250 U/mL muGM-CSF. After 48 hours of culture, the cells were pulsed with 0.5 μCi 3H-thymidine for 12 hours. The average 3H-thymidine incorporation among 3 control EML/Hyg clones and the average 3H-thymidine incorporation among 3 EML/Pax5A clones are shown and were found to be significantly different from each other using the heteroscededastic 1-tailed t-test (P < .001). (C) Three EML/Pax5A clones and 3 control EML/Hyg clones were split into SCF-containing growth media at 2.5 × 105 cells/mL and pulsed with3H-thymidine after 24 hours of culture for 17 hours. The average 3H-thymidine incorporation among 3 control EML/Hyg clones and the average 3H-thymidine incorporation among 3 EML/Pax5A clones are shown and were found to be significantly different from each other using the homoscededastic 2-tailed t-test (P < .004). (D) EML cells induced to differentiate as described in Fig 2A were replated at 5 × 105 cells/mL in 400 μL media containing 250 U/mL muGM-CSF. Two days later, the cells were stained with propidium iodide and then analyzed for apoptotic cells by flow cytometry. The average percentage of 3 EML/Hyg clones with subdiploid content of DNA and the average percentage of 3 EML/Pax5A clones with subdiploid content of DNA are shown and were found to be significantly different from each other using the homoscededastic 1-tailed t-test (P < .002).

BSAP/Pax5A-expressing EML cells fail to expand to myeloid growth factors. (A) EML cells were induced to differentiate into myeloid cells as described in Fig 2A and then replated in 1% methylcellulose with 250 U/mL muGM-CSF at 1.5 × 105 cells per 3.5-cm plate. On day 9, the colonies were aspirated and analyzed by flow cytometry to determine the mean number of CD11b+/Gr-1+ cells per colony. The average among 3 control EML/Hyg clones and the average among 3 BSAP/Pax5A-expressing EML clones are shown and were found to be significantly different from each other using the heteroscededastic 1-tailed t-test (P < .001). (B) EML cells induced to differentiate as described in Fig 2A were replated in suspension culture in the presence of 250 U/mL muGM-CSF. After 48 hours of culture, the cells were pulsed with 0.5 μCi 3H-thymidine for 12 hours. The average 3H-thymidine incorporation among 3 control EML/Hyg clones and the average 3H-thymidine incorporation among 3 EML/Pax5A clones are shown and were found to be significantly different from each other using the heteroscededastic 1-tailed t-test (P < .001). (C) Three EML/Pax5A clones and 3 control EML/Hyg clones were split into SCF-containing growth media at 2.5 × 105 cells/mL and pulsed with3H-thymidine after 24 hours of culture for 17 hours. The average 3H-thymidine incorporation among 3 control EML/Hyg clones and the average 3H-thymidine incorporation among 3 EML/Pax5A clones are shown and were found to be significantly different from each other using the homoscededastic 2-tailed t-test (P < .004). (D) EML cells induced to differentiate as described in Fig 2A were replated at 5 × 105 cells/mL in 400 μL media containing 250 U/mL muGM-CSF. Two days later, the cells were stained with propidium iodide and then analyzed for apoptotic cells by flow cytometry. The average percentage of 3 EML/Hyg clones with subdiploid content of DNA and the average percentage of 3 EML/Pax5A clones with subdiploid content of DNA are shown and were found to be significantly different from each other using the homoscededastic 1-tailed t-test (P < .002).

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