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. 2022 Feb 28;12:3259. doi: 10.1038/s41598-022-07228-x

Figure 4.

Figure 4

PD-1, PD-L1 and A2aR blockade enhances the toxicity of FLOT chemotherapy regimen demonstrated by a significant reduction in the percentage of viable cells and increase in late stage apoptotic/dead cells. OE33 cells and SK-GT-4 cells were treated with nivolumab (10 μg/ml), atezolizumab (10 μg/ml) or A2aR antagonist (3 μM) in the absence or presence of FLOT regimen for 48 h. Viability was determined by flow cytometry using annexin V propidium iodide assay. Necrotic cells ((A) AV−PI+), late stage apoptotic/dead cells ((B) AV+PI+), viable cells ((C) AV−PI−) and early stage apoptotic cells ((D) AV+PI−), dead were characterised. Representative dot plots shown for each treatment in OE33 cell line. Experiments repeated for n = 4 independent experimental repeats using singlet technical replicates. Kruskal–Wallis used for part (A–D) and Mann–Whitney test used for part (E,F), *p < 0.05, **p < 0.01 and ***p < 0.001.