Figure 8.
Figure 8. Stimulation of cell proliferation and chemotaxis by SDF-1α cleaved by CPN. (A) DW34 pre-B-cell proliferation in response to various concentrations (3 ng/mL-100 ng/mL) of SDF-1α, which was incubated for 10 minutes at room temperature with or without purified CPN (1:50 dilution) prior to testing. The results reflect the mean counts per minute (cpm) per culture (± SEM) of triplicate determinations; representative experiment of 4 performed. (B) BL41 cell migration across endothelial cell monolayers in the presence of SDF-1α (4 ng/mL-500 ng/mL) in the lower chamber of the transwell. SDF-1α was incubated for 10 minutes at room temperature with or without purified CPN (1:50 dilution) prior to testing. The results reflect the mean (± SEM) number of cells that have transmigrated to the lower chamber over a 4-hour incubation from 3 replicate wells per condition. Representative experiment of 5 performed. (C) DW34 pre-B-cell proliferation in response to full-length SDF-1α (4 ng/mL and 100 ng/mL) or SDF-1α lacking the carboxy-terminal lysine due to treatment with purified CPN (1:50 dilution, 10 minutes at room temperature), treatment with normal human plasma (1:10 dilution, 10 minutes at room temperature) or synthetic (SDF-1α 1-67). The results reflect the mean counts per minute (cpm) per culture (± SEM) of triplicate determinations; representative experiment of 3 performed. (D) BL41 cell migration across endothelial cell monolayers in the presence or absence of full-length SDF-1α (100 ng/mL) and CPN-treated SDF-1α (100 ng/mL) in the lower chamber of the transwell. In the presence of full-length SDF-1α (100 ng/mL) in the lower chamber of the transwell, BL41 cell transmigration was evaluated with full-length SDF-1α (100 ng/mL), SDF-1α (100 ng/mL) treated with purified CPN (1:50 dilution, 10 minutes at room temperature) or SDF-1α (100 ng/mL) treated with normal human serum (1:10 dilution, 10 minutes at room temperature) in the upper chamber of the transwell. The results reflect the mean (± SEM) number of cells that have transmigrated to the lower chamber over a 4-hour incubation from 5 replicate wells per condition. Representative experiment of 3 performed.

Stimulation of cell proliferation and chemotaxis by SDF-1α cleaved by CPN. (A) DW34 pre-B-cell proliferation in response to various concentrations (3 ng/mL-100 ng/mL) of SDF-1α, which was incubated for 10 minutes at room temperature with or without purified CPN (1:50 dilution) prior to testing. The results reflect the mean counts per minute (cpm) per culture (± SEM) of triplicate determinations; representative experiment of 4 performed. (B) BL41 cell migration across endothelial cell monolayers in the presence of SDF-1α (4 ng/mL-500 ng/mL) in the lower chamber of the transwell. SDF-1α was incubated for 10 minutes at room temperature with or without purified CPN (1:50 dilution) prior to testing. The results reflect the mean (± SEM) number of cells that have transmigrated to the lower chamber over a 4-hour incubation from 3 replicate wells per condition. Representative experiment of 5 performed. (C) DW34 pre-B-cell proliferation in response to full-length SDF-1α (4 ng/mL and 100 ng/mL) or SDF-1α lacking the carboxy-terminal lysine due to treatment with purified CPN (1:50 dilution, 10 minutes at room temperature), treatment with normal human plasma (1:10 dilution, 10 minutes at room temperature) or synthetic (SDF-1α 1-67). The results reflect the mean counts per minute (cpm) per culture (± SEM) of triplicate determinations; representative experiment of 3 performed. (D) BL41 cell migration across endothelial cell monolayers in the presence or absence of full-length SDF-1α (100 ng/mL) and CPN-treated SDF-1α (100 ng/mL) in the lower chamber of the transwell. In the presence of full-length SDF-1α (100 ng/mL) in the lower chamber of the transwell, BL41 cell transmigration was evaluated with full-length SDF-1α (100 ng/mL), SDF-1α (100 ng/mL) treated with purified CPN (1:50 dilution, 10 minutes at room temperature) or SDF-1α (100 ng/mL) treated with normal human serum (1:10 dilution, 10 minutes at room temperature) in the upper chamber of the transwell. The results reflect the mean (± SEM) number of cells that have transmigrated to the lower chamber over a 4-hour incubation from 5 replicate wells per condition. Representative experiment of 3 performed.

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