Figure 2.
Functionality of mbaIL6. (A) Jurkat cells (2 × 106/mL) transduced with either GFP alone (“Control”) or GFP plus mbaIL6 were cultured for 2 hours with 1 ng/mL human IL-6; IL-6 in the supernatant was measured by ELISA. Mean (±SD; n = 3) is shown. ****P < .0001. (B) Cultures were set up as in panel A; IL-6 levels were measured after the indicated time. The dashed curve is the fitted exponential decay curve. Mean (±SD; n = 3) is shown. (C) Cultures were set as in panel A but with various Jurkat cell concentrations; IL-6 was measured after 2 hours. Mean (±SD; n = 3) for each cell concentration is shown. **P < .01; ***P < .001; ****P < .0001. (D) Cultures were initiated with 0.1 × 106 cells/mL Jurkat cells; cell numbers increased to 0.2 × 106/mL after 24 hours, 0.4 × 106/mL after 48 hours, and 1.0 × 106/mL after 72 hours. Mean (±SD) for each time point is shown (n = 3). **P < .01; ****P < .0001. (E) Jurkat cells (2 × 106/mL) nontransduced (“wt”) or transduced with mbaIL6 were cultured for 2 hours with 1 ng/mL IL-6. U937 cells were exposed to the supernatant for 15 minutes at 37°C. Representative flow cytometry histograms show labeling of U937 cells with anti-STAT3 pY705. (F) Mean (±SD; n = 3) STAT3 phosphorylation in U937 cells. ***P < .001. (G) Varying concentrations of Jurkat cells transduced with mbaIL6 were cultured with 1 ng/mL IL-6 for 2 hours. The supernatant was added to 0.2 × 106 DS-1 cells, which were counted after 7 and 9 days of culture. Mean (±SD; n = 3) is shown. (H) Jurkat cells expressing mbaIL6 were cultured with IL-6 (5 ng/mL) for 2 hours; after washing, cells were cultured for another 2 hours and periodically labeled with anti–IL-6 PE. Flow cytometry dot plots show levels of IL-6 bound to mbaIL6 cells. Sequential data of 2 experiments are given in supplemental Figure 2C. (I) Jurkat cells (1 × 106/mL) expressing mbaIL6 were cultured with IL-6 (5 ng/mL) for 2 hours; after washing, cells were cultured for another 48 hours and periodically labeled with anti–IL-6 PE. Graph shows mean fluorescence intensity (MFI) of IL-6 (red) plotted together with cell count (blue). Mean (±SD; n = 3) is shown. (J) Supernatant from cultures shown in panel I was added to THP-1 cells for 15 minutes at 37°C. STAT3 phosphorylation was measured as in panel E. HL, half-life.

Functionality of mbaIL6. (A) Jurkat cells (2 × 106/mL) transduced with either GFP alone (“Control”) or GFP plus mbaIL6 were cultured for 2 hours with 1 ng/mL human IL-6; IL-6 in the supernatant was measured by ELISA. Mean (±SD; n = 3) is shown. ****P < .0001. (B) Cultures were set up as in panel A; IL-6 levels were measured after the indicated time. The dashed curve is the fitted exponential decay curve. Mean (±SD; n = 3) is shown. (C) Cultures were set as in panel A but with various Jurkat cell concentrations; IL-6 was measured after 2 hours. Mean (±SD; n = 3) for each cell concentration is shown. **P < .01; ***P < .001; ****P < .0001. (D) Cultures were initiated with 0.1 × 106 cells/mL Jurkat cells; cell numbers increased to 0.2 × 106/mL after 24 hours, 0.4 × 106/mL after 48 hours, and 1.0 × 106/mL after 72 hours. Mean (±SD) for each time point is shown (n = 3). **P < .01; ****P < .0001. (E) Jurkat cells (2 × 106/mL) nontransduced (“wt”) or transduced with mbaIL6 were cultured for 2 hours with 1 ng/mL IL-6. U937 cells were exposed to the supernatant for 15 minutes at 37°C. Representative flow cytometry histograms show labeling of U937 cells with anti-STAT3 pY705. (F) Mean (±SD; n = 3) STAT3 phosphorylation in U937 cells. ***P < .001. (G) Varying concentrations of Jurkat cells transduced with mbaIL6 were cultured with 1 ng/mL IL-6 for 2 hours. The supernatant was added to 0.2 × 106 DS-1 cells, which were counted after 7 and 9 days of culture. Mean (±SD; n = 3) is shown. (H) Jurkat cells expressing mbaIL6 were cultured with IL-6 (5 ng/mL) for 2 hours; after washing, cells were cultured for another 2 hours and periodically labeled with anti–IL-6 PE. Flow cytometry dot plots show levels of IL-6 bound to mbaIL6 cells. Sequential data of 2 experiments are given in supplemental Figure 2C. (I) Jurkat cells (1 × 106/mL) expressing mbaIL6 were cultured with IL-6 (5 ng/mL) for 2 hours; after washing, cells were cultured for another 48 hours and periodically labeled with anti–IL-6 PE. Graph shows mean fluorescence intensity (MFI) of IL-6 (red) plotted together with cell count (blue). Mean (±SD; n = 3) is shown. (J) Supernatant from cultures shown in panel I was added to THP-1 cells for 15 minutes at 37°C. STAT3 phosphorylation was measured as in panel E. HL, half-life.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal