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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: Inhibition of Stearoyl-CoA desaturase 1 reverts BRAF and MEK inhibition-induced selection of cancer stem cells in BRAF-mutated melanoma

Fig. 5

a) MUFA levels analysed by GS/MS in M14 and A375 BRAF/MEK plus MF438 treated cells; b) 12 Representative images of sphere formation of first generation taken on day 4. Scale bars: 50 μm. 13 Single-cell suspensions of M14, A375 and Mel 66 cell lines were seeded at 1000/well onto a 6-plate 14 ultra low attachment in sphere medium and treated with MF-438 alone or in combination with 15 BRAF/MEK inhibitors for 4 days; c) Sphere forming efficiency evaluated on A375, M14 and Mel 16 66 cell lines seeded at 1000/well onto a 96-plate ultra low attachment in sphere medium (3D). Cell 17 cultures treated with increasing concentrations of BRAF and MEK inhibitors (0.07-20 μM) 18 combined or not with MF-438 (0.07-50 μM). After 7 days of treatment the sphere-forming 19 efficiency of 3D cancer cells was compared to untreated cells; d) Proliferation assay performed on 20 2D and 3D cultures obtained from A375 and M14 cell lines exposed to MF-438 for 7 days; inset 21 shows the IC50 value calculated in 3D culture treated with BRAF/MEK and or BRAF/MEK plus 22 MF-438 (panel c) and IC50 3D vs 2D condition (panel d); e) Stemness markers (oct4, nanog, 23 jarir1b) analysed on M14 and A375 melanoma cells after BRAF/MEK plus MF-438 inhibitors by 24 qRT-PCR; f) Western blotting analysis of YAP/TAZ in M14 and Mel 66 spheroids treated with 25 BRAF, MEK or BRAF/MEK plus MF-438 for 96 hours; g) Immunofluorescence analyses of YAP/TAZ after BRAF/MEK inhibitors plus MF-438 performed on M14 and Mel 66 cell lines. 2 Scale bar 10mm; h) YAP/TAZ downstream target analysed after MF-438 combined with BRAF and 3 MEK inhibitors in A375, M14 and Mel 66

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