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. Author manuscript; available in PMC: 2020 Apr 1.
Published in final edited form as: Cell Immunol. 2019 Mar 19;338:32–42. doi: 10.1016/j.cellimm.2019.03.003

Figure 3. IDO1 protein synthesis and kynurenine activity in DCs treated with CTB-INS protein partially or completely purified from LPS.

Figure 3.

Panel (A), Human moDCs suspended in 2.0 mL of RPMI culture medium were treated with: 1) PBS as a negative control (NC), 2) LPS (1 μg/mL), 3) partially purified CTB-INS (10 μg/mL) isolated by nickel affinity column from E. coli BL-21 cells, containing residual levels of LPS, and 4) completely purified CTB-INS (10 μg/mL) treated with both Triton X-114 and Endotrap resin (T/E) to remove residual LPS. The levels of LPS were determined by Limulus amebocyte assay of all DC samples. Panel B, the DCs were cultured at 37°C for 24h and the amount of IDO1 protein synthesized in the DCs was identified by western blotting. Panel C, The activity of IDO1 synthesized in response to the different treatments was determined in all DC samples by kynurenine assay of the cell culture medium. The experimental data presented was from an individual donor representative of a total of three DC donors. The data for each measurement is expressed as the standard error of the mean of triplicate DC samples obtained from each donor. *p < 0.05, in comparison with the NC group.