Figure 8.
Figure 8. Fos expression in M1 cells partially abrogates the c-Myc block in myeloid differentiation and increases propensity of the cells to be induced for differentiation by IL-6. (A) Cell cycle analysis. Indicated cell types, either untreated or 3 days after treatment, were collected and subjected to FACS analysis as described in “Materials and methods.” Data shown are representative of at least 4 experiments. (B) Morphologic characteristics of myeloid differentiation. May-Grunwald-Giemsa-stained cytospin smears of indicated cell types, untreated or treated for 3 days, were assessed for morphologic markers associated with various stages of differentiation. At least 300 cells were scored to obtain the proportion of immature blast cells, cells at intermediate monocyte stages of differentiation, and mature macrophages. Data are an average (± SD) of at least 3 experiments that yielded similar results. Representative photomicrographs (original magnification, × 400) of cytospin smears are shown. (C) Expression of the macrophage differentiation markers ferritin and lysozyme. RNA was extracted from cells collected at indicated times following treatments. RNA was resolved on a 1% agarose formaldehyde gel and transferred to nylon membranes (Duralon) for Northern blot analysis. Blots were hybridized with a 32P-labeled ferritin probe, then stripped and reprobed with a 32P-labeled lysozyme probe as described in “Materials and methods.” Hybridization to an actin probe was used to ensure equal loading of RNA, as shown for Figure 2. (D-E) Increased propensity of MlFosER-myc cells to be induced for differentiation by suboptimal (1 ng/mL) IL-6. (D) Cell morphology following treatment with estrogen and suboptimal dose (ng/mL) of IL-6. May-Grunwald-Giemsa-stained cytospin smears of cells treated for 3 days with 1 ng/mL IL-6 plus estrogen were analyzed for morphologic differentiation characteristics. At least 300 cells were scored. (E) Lysozyme expression following treatment with estrogen plus 1 ng/mL IL-6. RNA blots (10 μg RNA/lane) were prepared with total RNA obtained at indicated time points after treatment and hybridized with a lysozyme cDNA probe labeled with 32P.

Fos expression in M1 cells partially abrogates the c-Myc block in myeloid differentiation and increases propensity of the cells to be induced for differentiation by IL-6. (A) Cell cycle analysis. Indicated cell types, either untreated or 3 days after treatment, were collected and subjected to FACS analysis as described in “Materials and methods.” Data shown are representative of at least 4 experiments. (B) Morphologic characteristics of myeloid differentiation. May-Grunwald-Giemsa-stained cytospin smears of indicated cell types, untreated or treated for 3 days, were assessed for morphologic markers associated with various stages of differentiation. At least 300 cells were scored to obtain the proportion of immature blast cells, cells at intermediate monocyte stages of differentiation, and mature macrophages. Data are an average (± SD) of at least 3 experiments that yielded similar results. Representative photomicrographs (original magnification, × 400) of cytospin smears are shown. (C) Expression of the macrophage differentiation markers ferritin and lysozyme. RNA was extracted from cells collected at indicated times following treatments. RNA was resolved on a 1% agarose formaldehyde gel and transferred to nylon membranes (Duralon) for Northern blot analysis. Blots were hybridized with a 32P-labeled ferritin probe, then stripped and reprobed with a 32P-labeled lysozyme probe as described in “Materials and methods.” Hybridization to an actin probe was used to ensure equal loading of RNA, as shown for Figure 2. (D-E) Increased propensity of MlFosER-myc cells to be induced for differentiation by suboptimal (1 ng/mL) IL-6. (D) Cell morphology following treatment with estrogen and suboptimal dose (ng/mL) of IL-6. May-Grunwald-Giemsa-stained cytospin smears of cells treated for 3 days with 1 ng/mL IL-6 plus estrogen were analyzed for morphologic differentiation characteristics. At least 300 cells were scored. (E) Lysozyme expression following treatment with estrogen plus 1 ng/mL IL-6. RNA blots (10 μg RNA/lane) were prepared with total RNA obtained at indicated time points after treatment and hybridized with a lysozyme cDNA probe labeled with 32P.

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