Skip to main content
. Author manuscript; available in PMC: 2022 Jul 1.
Published in final edited form as: Mol Cancer Ther. 2021 Nov 4;21(1):170–183. doi: 10.1158/1535-7163.MCT-21-0194

Figure 4. Vertical inhibition of MAPK pathway synergistically inhibits ERK activity and H/NRAS Q61X-mutant FN-RMS cells growth.

Figure 4.

(A) Effect of vertical targeting of MAPK pathway on ERK activity as measured by phosphorylated ERK (pERK, Thr202/Tyr204)) and phosphorylated p90RSK (pRSK, Thr573). Apoptosis marker is also shown (cleaved PARP). Cells were treated with DMSO (control), 2 nM trametinib, 100 nM ERKi LY3214996, 100 nM pan-RAFi LY3009120 or combinations for 72 h and cell lysates analyzed by western blots. (B) Growth curves of RMS cells following treatment with DMSO, trametinib, ERKi LY3214996, pan-RAFi LY3009120 or combinations. Cells were plated in 96 well plates and next day treated as in A. Percent confluency was determined by imaging with Incucyte every 2 h for 168 h. (C) Crystal violet staining images of RAS-mutant RMS cell lines and one RAS wild-type (Rh18) upon treatment with trametinib, ERKi LY3214996, pan-RAFi LY3009120 or combinations. Cells were plated at low density in 12-well plates and next day treated with DMSO or increasing concentrations of the indicated drugs/combinations. After two weeks, cell growth was visualized with crystal violet staining.