IL35 (Interleukin 35) is an IL-12 family cytokine, which is a dimeric protein composed of IL-12α and IL-27β chains. IL35 is thought to mediate the immune inhibitory function of regulatory T cells and has been proven to promotes pancreas cancer growth through enhancement of proliferation and inhibition of apoptosis. Thus, proliferation assay of IL35 was conducted using PANC-1 cells. Briefly,
PANC-1 cells were seeded into triplicate wells of 96-well plates at a density of 2,000 cells/well and allowed to attach overnight, then the medium was replaced with serum-free standard DMEM?prior to the addition of various concentrations of recombinant human IL35. After incubated for 48h,?cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10μL of CCK-8 solution was added to each well of the plate, then the absorbance at 450nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37°C. Proliferation of PANC-1 cells after incubation with IIL35 for 48h observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8 ) assay after incubation with human recombinant IL35 for 48h. The result was shown in Figure 2. It was obvious that IL35 significantly increased cell viability of PANC-1 cells.