Figure 2.

GBM proliferation, migration, and invasion, promoted by IL-1β stimulation, was mitigated after treatment with anakinra (Ana). T98G cells were stimulated with IL-1β in the presence or absence of anakinra. (A) Quantification of Ki-67-positive T98G cells by flow cytometry (n = 5, p = 0.027). One representative histogram is shown. (B) Analysis of chemotaxis time-lapse microscopy by single-cell tracking. IL-1β-stimulated T98G cells were incubated with FCS as a chemotactic stimulus (right reservoir). At least 40 cells were tracked. One representative example of four independent experiments is shown (left panel, n = 4). Analysis of forward migration index (FMI, coordinate of a cell in the indicated direction (x-axis) divided by the accumulated distance of its paths, representing efficiency of forward migration) (right panel, n = 4, p = 0.048). (C) Analysis of tumor cell invasion by transwell invasion assay. Stimulated T98G cells were seeded in cell culture inserts and allowed to migrate towards FCS as a chemotactic stimulus. Optical density of invasive T98G represents the amount of transmigrated cells (n = 3, p < 0.05). (D) 2D migration assay of GBM cells, stimulated with IL-1β in the presence or absence of anakinra at start and after 12h. Lines mark the initially cell-free area. A typical example of six experiments is shown (n = 6). * p < 0.05.