EFTX-D1 inhibits mutant
KRAS-driven cancer cell growth. (a) Flow
cytometry cell cycle analysis of H358 (KRAS G12C), H441 (KRAS G12V),
and A549 (KRAS G12S) lung cancer cells 72 h post transient transfection
with NC, Seq2, or EFTX-D1 siR at 20 nM. (b) Western blot analysis
of KRAS, pERK, and total ERK expression in H358 (KRAS G12C), H441
(KRAS G12V), and A549 (KRAS G12S) lung cancer cells transiently transfected
with NC, Seq2, or EFTX-D1 siR at 20 nM. Cells were analyzed 36 h post
transfection. (c) 2-D proliferation of H358, H441, and A549 lung cancer
cells transiently transfected with NC, Seq2, or EFTX-D1 siR at 20
nM. Cells were quantified after 7 days of growth. Samples were run
in at least triplicate, and cell counts are normalized to NC. (d)
3-D growth of A549 and H441 lung cancer cells embedded in Matrigel
24 h after transient transfection with NC, Seq2, or EFTX-D1 siR at
20 nM. Cells were imaged and quantified using OrganoSeg software after
7 days of growth, and conditions were run in at least triplicate;
scale bar 400 μm. Statistical significance was measured by a
one-way ANOVA with a Tukey’s multiple comparisons test; p-values are indicated as **p < 0.01,
***p < 0.001. Error bars represent standard error
of the mean.