Fig 2: Drug treatments increase the correlations between the activity of pathways.A Scatter plots of the activity scores of p38 and p53 reporters in the A375 signalome cell line before and at multiple time points after treatment with 122 active drugs. Each dot represents a different drug. Pearson’s r is depicted for each of the time points. B Pearson correlation coefficients were calculated for each pair of pathways in the A375 signalome cell line before and at different time points after treatment with 122 active drugs. Each datapoint represents a correlation value for one given pair of pathways over all 122 active drugs. The correlation between the activity score of p38 and p53 pathways is marked by a red circle. C–H Locally weighted smoothing (Lowess) regression of all reporters in each of the two signaling states is shown for six representative drug treatments, each associated with different drug targets. While three of the drugs drive the p53-signaling state (C–E), the other three drugs drive the p38 signaling state (F–H). The bold (red and blue) lines and the gray sleeves represent mean values and + /- SEM respectively. Source data are provided with this paper.

Fig 3: mFGF21 translates into functional protein in human adipocytes(A) Representative immunostaining and ELISA analysis of human FGF21 in cells transfected with control LNP and mFGF21. (B–D) Phosphorylation of ERK in adipocytes treated with recFGF21 and mmFGF21. (E) Glucose uptake in adipocytes stimulated by recFGF21 or mFGF21. (F) mFGF21 protein in conditioned media of transfected adipocytes. (G) FGF21 down-stream genes expression in human adipocytes treated with recFGF21 or mFGF21. (H) Injection site luciferase quantification performed at 6, 24, and 48 h after injection. (I) Representative image of RNA distribution at the injection site (brown signal; RNAscope) was taken at 6 h after single s.c. administration of mmLUC 0.3 mg/kg. Figures show mean ± standard error of the mean. n = 4–8, *p < 0.05, **p < 0.01, and ***p < 0.001 versus PBS, #p < 0.05, ##p < 0.01, and ###p < 0.001 versus LNP. Ordinary one-way ANOVA was performed followed by Sidak’s multiple comparison test.

Fig 4: Cathepsin S (CatS) deletion improved targeted molecule changes in the injured arteries of the stressed mice. A through C, Representative images and combined quantitative data for the levels of targeted molecule proteins. The results are mean±SEM (n=5–7). AKT indicates protein kinase B; ATR1, angiotensin II receptor 1; CatS+/+, CatS wild type; CatS-/-, CatS deficient; Erk, extracellular signal-regulated kinase; GSK, phosphoglycogene synthesis kinase; mTOR, mammalian target of rapamycin; NS, no significance; p-AKT, phosphorylated AKT; p-Erk, phosphorylated Erk; p-GSK, phosphorylated GSK; p-mTOR, phosphorylated mTOR. ***P<0.001 vs CatS+/+ by 1-way ANOVA, followed by Tukey post hoc tests or Student t test.

Fig 5: Stress altered the levels of targeted protein in the injured arteries. A through C, Representative images and combined quantitative data for the levels of targeted proteins. Results are mean±SEM (n=5–7). AKT indicates protein kinase B; ATR1, angiotensin II receptor 1; CatS+/+, cathepsin S wild type; Erk, extracellular signal-regulated kinase; GSK, phosphoglycogene synthesis kinase; mTOR, mammalian target of rapamycin; NS, no significance; p-AKT, phosphorylated AKT; p-Erk, phosphorylated Erk; p-mTOR, phosphorylated mTOR. ***P<0.001 vs the relative controls by 1-way ANOVA, followed by Tukey post hoc tests.