Figure 5.
Figure 5. Analysis of differentiation-associated characteristics of M1, M1myc, M1Egr-1, and M1Egr-1/myc cells. Cells were seeded at 0.05 × 106 cells/mL with or without IL-6 (50 ng/mL). (A) After 3 days cells were harvested and cell morphology was determined, as described in Figure 3, and “Materials and methods.” Results are the average of 3 independent experiments yielding similar results with an SD up to 10%. Using mature cells as a measure of cell differentiation, the differences between M1myc and each of the other 3 cell lines used were P < .01 (significant), between M1 and M1Egr-1/myc P > 0.1 (not significant), and between either M1 or M1Egr-1/myc and M1Egr-1 P < .05 (significant). (B) Photomicrographs of M1, M1myc, M1Egr1, and M1Egr1-myc cells from May-Grünwald-Giemsa–stained cytospin smears untreated and after 3 days of treatment with either 1 ng/mL or 50 ng/ml IL-6. Slides were analyzed and photographed (original magnification, × 400). Images were cpatured with a Zeiss Axioplan microscope using a 40×/0.75 NA Neofluar lens and a SenSys camera (Roper Scientific, Tucson, AZ) and SmartCapture 2 software (Digital Scientific, Cambridge, United Kingdom). (C) Expression of ferritin and lysozyme mRNA was determined by Northern blot analysis using of total RNA as described in “Materials and methods.” This is a representative experiment that was carried out 3 times. (D) F4-80 expression was determined using FITC-conjugated anti–F4-80 primary antibody and flow cytometry, as described in Figure 3 and “Materials and methods.” (E) Photomicrographs of phagocytosis analysis showing cells phagocytosing latex beads. Cells were seeded in the absence or presence of IL-6 and incubated with or without latex beads as described in “Materials and methods.” Images were captured using a Nikon dissectin microscope (SMZ-U, zoom 1:10) and an ED Plan APO lense (Nikon, Surrey, United Kingdom) and a Nikon CoolPix 990 camera and were processed with Adobe Photoshop (Adobe Systems, San Jose, CA). Photomicrographs are representative of at least 30 fields examined per cell line. Results are shown for M1Egr-1/myc6. Similar data were obtained with M1Egr-1/myc7.

Analysis of differentiation-associated characteristics of M1, M1myc, M1Egr-1, and M1Egr-1/myc cells. Cells were seeded at 0.05 × 106 cells/mL with or without IL-6 (50 ng/mL). (A) After 3 days cells were harvested and cell morphology was determined, as described in Figure 3, and “Materials and methods.” Results are the average of 3 independent experiments yielding similar results with an SD up to 10%. Using mature cells as a measure of cell differentiation, the differences between M1myc and each of the other 3 cell lines used were P < .01 (significant), between M1 and M1Egr-1/myc P > 0.1 (not significant), and between either M1 or M1Egr-1/myc and M1Egr-1 P < .05 (significant). (B) Photomicrographs of M1, M1myc, M1Egr1, and M1Egr1-myc cells from May-Grünwald-Giemsa–stained cytospin smears untreated and after 3 days of treatment with either 1 ng/mL or 50 ng/ml IL-6. Slides were analyzed and photographed (original magnification, × 400). Images were cpatured with a Zeiss Axioplan microscope using a 40×/0.75 NA Neofluar lens and a SenSys camera (Roper Scientific, Tucson, AZ) and SmartCapture 2 software (Digital Scientific, Cambridge, United Kingdom). (C) Expression of ferritin and lysozyme mRNA was determined by Northern blot analysis using of total RNA as described in “Materials and methods.” This is a representative experiment that was carried out 3 times. (D) F4-80 expression was determined using FITC-conjugated anti–F4-80 primary antibody and flow cytometry, as described in Figure 3 and “Materials and methods.” (E) Photomicrographs of phagocytosis analysis showing cells phagocytosing latex beads. Cells were seeded in the absence or presence of IL-6 and incubated with or without latex beads as described in “Materials and methods.” Images were captured using a Nikon dissectin microscope (SMZ-U, zoom 1:10) and an ED Plan APO lense (Nikon, Surrey, United Kingdom) and a Nikon CoolPix 990 camera and were processed with Adobe Photoshop (Adobe Systems, San Jose, CA). Photomicrographs are representative of at least 30 fields examined per cell line. Results are shown for M1Egr-1/myc6. Similar data were obtained with M1Egr-1/myc7.

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