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. 2016 Oct 3;8(5):8807–8817. doi: 10.18632/oncotarget.12410

Figure 1. Confirmation of PD-L1 expression as a target for NIR-PIT in H441 cells, and evaluation of in vitro NIR-PIT.

Figure 1

A. Validation of avelumab-IR700 by SDS-PAGE (left: Colloidal Blue staining, right: fluorescence). Diluted avelumab was used as a control. B. Expression of PD-L1 in H441 cells was examined with FACS. After 6 h of avelumab-IR700 incubation, H441 cells showed high fluorescence signal. C. Expression of PD-L1 in H441 cell was estimated by expression of EGFR in A431 cell using FACS. Mean fluorescence of H441 cell with avelumab-IR700 was 16.2, on the other hand the mean fluorescence of A431 cell with panitumumab-IR700 was 177.8. D. Differential interference contrast (DIC) and fluorescence microscopy images of H441 cells after incubation with avelumab-IR700 for 6 h. High fluorescence intensities were shown in H441 cells. Necrotic cell death was observed upon excitation with NIR light (after 15min). Scale bars = 20 μm. E. Membrane damage of cells induced by PIT was measured with the dead cell count using PI staining, which increased in a light dose dependent manner (n = 5, *p < 0.01, vs. untreated control, by Student's t test).