American Society of Hematology

Figure 2.

$Figure 2. CXCL7 gene expression and protein secretion in CD14+ cells. (A) cDNA was prepared from CD14+ cells after 5 days of culture in control media (RPMI with 10% FCS), media conditioned by HS5 cells, media conditioned by primary long-term cultures (LTC CMs), or media conditioned by human foreskin fibroblasts (HFFs). CXCL7 gene expression was measured by Syber Green real-time PCR as in “Materials and methods”. Data represent mean ± SE of 2 experiments. Both HS5 CM and LTC CM differ from control and HFF CM by P = .004 and .003, respectively, using Student t test. (B) To measure the time course of CXCL7 up-regulation, cDNA was prepared from CD14+ cells cultured in HS5 CM for up to 8 days. CXCL7 gene expression, measured by real-time PCR, is shown on the y-axis on the right. The media isolated from the same cultures were assayed for CXCL7 protein content using ELISA, shown on the y-axis to the left. (C) Conditioned media obtained from HS5 cultures was fractionated by filtration with a 10-kDa cutoff. CD14+ cells were cultured for 3 days in control media, HS5 CM, the smaller than 10-kDa fraction, the larger than 10-kDa fraction, and a combination of both fractions. Results are shown as mean of at least 2 samples ± SE.$

CXCL7 gene expression and protein secretion in CD14⁺ cells. (A) cDNA was prepared from CD14⁺ cells after 5 days of culture in control media (RPMI with 10% FCS), media conditioned by HS5 cells, media conditioned by primary long-term cultures (LTC CMs), or media conditioned by human foreskin fibroblasts (HFFs). CXCL7 gene expression was measured by Syber Green real-time PCR as in “Materials and methods”. Data represent mean ± SE of 2 experiments. Both HS5 CM and LTC CM differ from control and HFF CM by P = .004 and .003, respectively, using Student t test. (B) To measure the time course of CXCL7 up-regulation, cDNA was prepared from CD14⁺ cells cultured in HS5 CM for up to 8 days. CXCL7 gene expression, measured by real-time PCR, is shown on the y-axis on the right. The media isolated from the same cultures were assayed for CXCL7 protein content using ELISA, shown on the y-axis to the left. (C) Conditioned media obtained from HS5 cultures was fractionated by filtration with a 10-kDa cutoff. CD14⁺ cells were cultured for 3 days in control media, HS5 CM, the smaller than 10-kDa fraction, the larger than 10-kDa fraction, and a combination of both fractions. Results are shown as mean of at least 2 samples ± SE.

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