1. Adhesion receptor expression by hematopoietic cell lines and murine progenitors 
Pamela S Becker, Susan K Nilsson, Zhifang Li, Virla M Berrios, Mark S Dooner, Cathleen L Cooper, Chung-cheng Hsieh, Peter J Quesenberry 
Experimental Hematology 
Volume 27, Issue 3, Pages (March 1999)
DOI: /S X(98)00037-X

2. Figure 1
Northern blot analysis: α4 integrin is downregulated on HL-60 cells after incubation with a combination of cytokines. HL-60 cells were incubated for 48 hours without cytokines (0), with individual cytokines (IL-3, IL-6, IL-11, or SCF), or a combination of all 4 cytokines (cocktail). A probe to α4 integrin was used for hybridization of the Northern blot. A probe to the ribosomal 28S subunit was used as a control, and the amount of α4 was normalized to expression of the 28S ribosomal RNA
Experimental Hematology  , DOI: ( /S X(98)00037-X)

3. Figure 2
Quantitative RT-PCR for α4 integrin on synchronized FDC-P1 cells. The dashed curve indicates the counts per minute of 3H-thymidine incorporation into DNA. The peak at about 12 hours represents S phase. The amount of mRNA for α4 integrin is represented by the solid curve, as determined by quantitative RT-PCR, and indicated in moles × 10−22. The data depicted are representative of three separate experiments
Experimental Hematology  , DOI: ( /S X(98)00037-X)

4. Figure 3
Surface expression of α4 on synchronized FDC-P1 cells. The dashed curve indicates the counts per minute of 3H-thymidine incorporation into DNA, in separate experiments from those described in Figure 2. The peak at about 12 hours represents S phase. The solid curve indicates the percent of cells expressing α4 integrin by flow cytometry analysis. The Y axis indicates the percent of cells expressing α4 integrin. The error bars represent standard deviation
Experimental Hematology  , DOI: ( /S X(98)00037-X)

5. Figure 4
RT-PCR products derived from Lin−Sca+ and Lin−Hoechst 33342lo/rhodamine 123lo cells. The left panel shows the RT-PCR products derived from RNA isolated from Lin−Sca+ cells, and the right panel, those derived from RNA isolated from Lin−Hoechst33342lo/rhodamine 123lo cells, utilizing primers unique for each of the α chains, α1- α6. The Lin−Sca+ cells express α1, α3, α5, and α6, whereas the Lin−Hoechst33342lo/rhodamine 123lo cells express α3, α5, and α6
Experimental Hematology  , DOI: ( /S X(98)00037-X)

6. Figure 5
Nucleotide sequences from integrin RT-PCR products derived from Lin−Sca+ cells. Partial nucleotide sequences were obtained by RT-PCR from RNA derived from the sorted progenitor cell populations. The upper line represents the sequence of the PCR products and the lower line, the sequence of the indicated receptor isoform obtained from Genbank. The sequences confirmed identity with known murine isoforms, including α1, α3A, α5, and α6B
Experimental Hematology  , DOI: ( /S X(98)00037-X)

7. Figure 6
Adhesion receptor expression on Lin−Sca+ cells by immunofluorescence. One example of a highly expressed receptor, CD44, is seen in the upper panels, and a receptor not expressed, α2, is seen in the lower panels. DAPI (blue) was used as a counterstain for all cell nuclei. The anti-Sca antibody was PE-labelled (red), the anti-receptor antibodies were visualized with FITC-anti-rat IgG (green). A dual filter was used to visualize the simultaneous expression of Sca and the adhesion receptor. Note that the FITC stain is clearly visible for CD44, and negative for α2
Experimental Hematology  , DOI: ( /S X(98)00037-X)

8. Figure 7
Flow cytometry analysis of adhesion receptor expression. Cytokine incubation of murine bone marrow cells from which the Lin−Sca+ cells were isolated. Murine bone marrow cells were incubated with the four cytokines, IL-3, IL-6, IL-11, and SCF as described in Methods, for 0, 24, and 48 hours at 37°C in Teflon bottles. The cells were then lineage depleted, labeled with PE-Sca+ and the Lin−Sca+ cells isolated by FACS. These cells were then analyzed by flow cytometry for adhesion receptor expression, and exhibit the indicated percents of positive cells. The expression of α1, α4, and β1 declined by 48 hours, whereas the expression of L-selectin increased at 24 hours, and the expression of PECAM-1, α5, and αL was increased at both 24 and 48 hours. The error bars represent standard errors of the mean; p < for all receptors comparing 24 or 48 hours to 0 hours except β1, for which p = 0.07 comparing 24 hours to 0 hours, and p < comparing 48 hours to 0 hours
Experimental Hematology  , DOI: ( /S X(98)00037-X)

9. Figure 8
Cytokine incubation reduces functional adhesion to extracellular matrix (ECM) components and stroma. The shaded bars indicate 0 hours, and the solid bars 48 hours of cytokine incubation prior to the adhesion assay. Lin−Sca+ cells were incubated on ECM components or stroma for 2 hours at 37°C in culture slides. The slides were then washed three times with PBS and the adherent cells counted as described in Methods. The y axis indicates the average number of cells per high powered field. mFn = mouse fibronectin; hRn = human Retronectin; mCol I = mouse collagen I; Ln = mouse laminin; TC-1 = stroma cell line; Dex = Dexter stroma after 4 weeks of culture
Experimental Hematology  , DOI: ( /S X(98)00037-X)