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. Author manuscript; available in PMC: 2024 Mar 1.
Published in final edited form as: Mol Cancer Res. 2023 Sep 1;21(9):867–880. doi: 10.1158/1541-7786.MCR-22-1031

Figure 1. BRAFi-resistant, but not sensitive, cell lines exhibit a pro-invasive phenotype and secretome in response to BRAF inhibition.

Figure 1.

A) BRAF-V600E cell lines that are sensitive (KTC1, CUTC5), have acquired resistance (KTC1-VEMR), or intrinsic resistance (T238, 8505C, CUTC60) to BRAF inhibition were treated with BRAFi for 24 hrs then plated in Matrigel-coated Boyden chambers for 24 hrs. Invading cells were stained with DAPI and counted using ImageJ or the Cytation 5 Cell Imaging Multi-Mode Reader. Representative images shown. B) CM was collected from sensitive (SENS-CM) or resistant (RES-CM) cell lines pre-treated with vehicle or BRAFi. BRAFi-sensitive or –resistant cell lines were treated with CM from indicated cell lines for 24 hrs then plated in Matrigel-coated Boyden chambers for 24 hrs, stained with DAPI and counted using ImageJ or the Cytation 5 Cell Imaging Multi-Mode reader. A-B) Results displayed as mean normalized to DMSO treated control. C) CUTC5 or CUTC60 cells were plated in ImageLock 96-well plates, allowed to adhere overnight, scratched with a WoundMaker, then overlaid with Matrigel supplemented with CM from vehicle- or BRAFi-pretreated CUTC60 cells. Percent wound confluence was measured every 4 hrs for 72 hrs using IncuCyte® ZOOM live-cell imaging. Mean area under the curve of percent wound confluence over time +/− SEM. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns = not significant. BRAFi: 100 nM dabrafenib, SENS-CM: conditioned media collected from BRAFi-sensitive cells, RES-CM: conditioned media collected from BRAFi-resistant cells.