Figure 3.

Targeting PLK1 elicited ICD in vitro and in vivo. A-E. LLC cells were treated for 24 h with indicated concentrations of BI2536. A. Cell viability was measured by CCK-8 assay. B. Immunofluorescence imaging of CRT translocation (green), phalloidin (red, cellular cytoskeleton) and DAPI (blue, nuclei). Scale bars, 60 µm. C-D. The data of CRT expression by Flow cytometry. E. Release of ATP in cell culture supernatants. F-J. LLC cells treated in vitro with BI2536 or by freezing-thawing were inoculated into right flank of immunocompetent C57BL/6 mice. Eight days after inoculation, the mice were challenged with live LLC cells in the opposite flank, as indicated in treatment scheme (G). The survival condition and tumor size were monitored every 2-3 days. F. Percentage of tumor-free mice in each group (n=8). H-J. Growth of challenged tumors in an established immunization model.