Figure 2. Cell proliferation of TF-1 cells after stimulated with recombinant dog IL6.

Interleukin 6 (IL-6) is an interleukin that acts as both a pro-inflammatory cytokine and an anti-inflammatory myokine. Interleukin 6 is secreted by T cells and macrophages to stimulate immune response and also plays a role in fighting infection. It supports the growth of B cells and is antagonistic to regulatory T cells. To test the effect of IL6 on cell proliferation, TF-1 cells were seeded into triplicate wells of 96-well plates at a density of 20,000 cells/well with various concentrations of recombinant dog IL6. After incubated for 72 hours,?cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8(CCK-8). Briefly, 10 μl of CCK-8 solution was added to each well of the plate, then the absorbance at 450 nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37℃. Proliferation of TF-1 cells after incubation with IL6 for 72 hours observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8 ) assay after incubation with recombinant dog IL6 for 72 hours. The result was shown in Figure 2. It was obvious that recombinant dog IL6 significantly increased cell viability of TF-1 cells, the EC50 was 0.04-0.08 ug/ml.