Fig. 4: Combined inhibition of KRAS^G12C and mTORC1 blocks Cyclin D1 expression at the transcriptional and translational levels.

a H1373 cells were treated with 1 nM RMC-6272, 100 nM RM-018, or the combination for 24 h. Cell cycle states were detected with EdU-DAPI-based flow cytometry. b H1373 and H2122 cells were treated with either 100 nM RM-018, 1 nM RMC-6272, or the combination for the indicated times, and lysates were analyzed by immunoblot. c Schema of the flow for the methionine starvation experiment in (d). d H1373 cells were cultured in methionine starved medium for 24 h, followed by adding back complete media (CM) alone, or with either 1 nM RMC-6272 or 50 µg/ml cycloheximide (CHX) for the indicated times, and lysates were analyzed by immunoblot. e Cyclin D1 mRNA levels were determined by quantitative PCR in H1373, HOP62, H2030, and H2122 cells following 1 nM RMC-6272, 100 nM RM-018, or the combination for 24 h. The data shown represent the means ± SD of n = 3 experimental replicates. P values are shown and were calculated by one-way ANOVA with post-hoc Tukey’s test.