Figure 6.

ROCK-Myosin II Inhibition Induces Lethal ROS, DNA Damage, and Cell-Cycle Arrest

(A) GSEA of ROS/oxidative stress-related gene signatures in MAPKi-resistant versus sensitive cell lines (group 1) from (Song et al., 2017). Dashed line indicates statistical significance.

(B) The 10 most enriched canonical pathways in downregulated genes in MAPKi-resistant cell lines (group 1) from (Song et al., 2017).

(C) Left, ROS levels in A375 (s) and A375/PLX/R (R) cells after treatment (n = 6). Right, quantification of p-H2A.X immunoblots (n = 7).

(D and E) Cell-cycle analysis after treatment (n = 3–4). Sensitive (s)-resistant (R) pairs (D, left A375; right WM983A). A375/PLX/R (E); G, GSK269962A; B, blebbistatin.

(F) p-STAT3 levels after treatment (n = 3 patient no. 35, WM793B; n = 4 A375/PLX/R; n = 5 patients no. 2 and 58).

(G) Mcl-1 levels of A375/PLX/R cells after treatment (n = 3).

(H and I) Percentage of dead cells by annexin V/PI staining of A375/PLX/R (H), patient no. 2 and no. 62T3 (I) cells after a 3-day treatment (n = 4 A375/PLX/R; n = 5 patient no. 2; n = 4 patient no. 62T3).

(C, D, and F–I) ROCKi GSK269962A; (E, H, and I) BRAFi PLX4720.

(C–G) 24 h treatment.

(C and F–I) Mean ± SEM and individual data points (circle). Asterisks in (D and E) are statistical significance in multinucleated cells. p values by one-way ANOVA with Tukey's (D–F, H, and I) or Benjamini, Krieger, and Yekutieli correction (C); unpaired t test with Welch's correction (G), ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001; n.s., not significant. See also Figure S6.