Fig. 6
From: Modulating multi-functional ERK complexes by covalent targeting of a recruitment site in vivo
BI-78D3 overcomes acquired resistance to BRAF inhibition in melanoma cell lines. a–d BI-78D3 is equally potent in PLX4032-sensitive (-S) or -resistant (-R), BRAF-mutant melanoma cell lines. Cells were treated with either PLX4032 or BI-78D3 as shown and incubated for 72 h before analysis using an MTS assay (Error bars: ±SD (n = 3)). e, f BI-78D3 inhibits the anchorage-dependent growth of PLX4032-sensitive and -resistant melanoma cell lines (experiments were repeated at least two times). g, h BI-78D3 induces caspase-3/7 activity in 451 Lu-S and 451 Lu-R cell lines. Cells were analyzed using an IncuCyte caspase-3/7 reagent and the IncuCyte® ZOOM at the indicated time points. (IncuCyte imaging. Error bars: SEM (n = 3)). i Western blot analysis to evaluate the suppression of EGF-stimulated cell signaling in melanoma cell lines by BI-78D3. Cells were serum starved overnight then treated with BI-78D3 for 1 h in serum-free media, followed by inhibitor washout for 2 h in the same media. Cells were then stimulated with EGF for 30 min in full media. Images have been cropped for presentation; the uncropped images are shown in Supplementary Fig. 27
