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. 2022 Jan 12;13(1):54. doi: 10.1038/s41419-022-04502-8

Fig. 1. Conditioning to IZI5 renders MM cells resistant to TRAIL-receptor-agonist-induced apoptosis and causes expansion of 3D spheroids.

Fig. 1

A Apoptosis induction was monitored 16 h after stimulation of parental (p) and IZI5-conditioned (c) BRAF-mutated (mutBRAF) A375 and Malme3M as well as NRAS-mutated (mutNRAS) WM1346 and WM1366 MM cells with IZI50 using a Cell Death Detection ELISA (CDDE) (n = 3: ***p ≤ 0.001). B Processing of caspase-8, caspase-3, and PARP was assessed by Western-blot analysis with GAPDH as loading control. One representative out of three independently performed experiments is shown. C GFP-expressing green fluorescent spheroids consisting of parental (p) and IZI5-conditioned (c) melanoma cells were individually embedded into 3D dextran-based gel-matrices and stimulated with IZI50 every other day over 8 days (scale bar = 200 µm). At day eight cell death was visualized by addition of 6.7 µg/ml PI. Confocal images of individual spheroids at day one and eight and the quantification of tumor volume (green) versus tumor death (red) at day eight are shown (n = 3: *p ≤ 0.05; ***p ≤ 0.001; n.s. = not significant).