Fig. 5: Interleukin (IL)−6-STAT3-AKT induces the proliferation of bronchial smooth muscle cells (SMC) via FoxO1. | Nature Communications

Fig. 5: Interleukin (IL)−6-STAT3-AKT induces the proliferation of bronchial smooth muscle cells (SMC) via FoxO1.

From: Maternal and perinatal obesity induce bronchial obstruction and pulmonary hypertension via IL-6-FoxO1-axis in later life

Fig. 5

A Immunoblot showing phosphorylated STAT3 (pSTAT3), and total STAT3 in primary murine bronchial smooth muscle cells (bSMC) from wild-type mice (WT). B Nuclear and cytoplasmatic phosphorylated FoxO1 (pFoxO1) fraction in % of total cell pFoxO1 per bSMC; pFoxO1 (green) and DAPI (blue, nucleus). C Nuclear and cytoplasmatic total FoxO1 fraction in % of total cell FoxO1 per bSMC. D Total FoxO1 in WT-bSMC. E Nuclear and cytoplasmatic pFoxO1 fraction in % of total cell pFoxO1 and total nuclear FoxO1 per cell in WT-bSMC. F Proliferation (cell count) or viability (MTT assay) in WT-bSMC. G Viability (MTT assay), proliferation (BrdU), and apoptosis (active caspase3/7 assay) in WT-bSMC. H Immunoblot showing phosphorylated AKT (pAKT), and total AKT in WT-bSMC. I Nuclear and cytoplasmatic pFoxO1 fraction in % of total cell pFoxO1 per WT-bSMC. J Immunoblots showing pSTAT3, total STAT3, pAKT, and total AKT in WT-bSMC. K Nuclear and cytoplasmatic pFoxO1 fraction in % of total cell pFoxO1 per WT-bSMC. L, M Nuclear and cytoplasmatic total FoxO1 fraction in % per bSMC from mice with inducible activation of FoxO1 (FoxO1ADA mice). N Immunoblot of WT-bSMC and IL-6−/−bSMC showing pSTAT3 and total STAT3. O Nuclear and cytoplasmatic total FoxO1 fraction in % of total cell FoxO1 per bSMC. P Viability (MTT assay) and proliferation (BrdU). Q–S immunoblot for pSTAT3 and STAT3 in WT-bSMC (Q). Viability (MTT assay), proliferation (cell count), and pSTAT3 per cell using immunocytochemistry in bSMC from mice with a specific ablation of FoxO1 in SMCs (SMCFoxO1 KO) and WT mice (R, S). For immunoblots, β-actin was used as a loading control; densitometric summary is displayed. Interleukin-6 and soluble IL-6 receptor (IL-6 + R), Stattic (STAT3 inhibitor), insulin, AKT Inhibitor (AKT-Inh.), Paclitaxel, and FoxO1 Inhibitor (FoxO1Inh). All bSMC are isolated from 21-day-old mice. Data are shown as mean ± standard error of the mean. A–S n = 3–9 biological independent samples or experiments per group; (repeated measure) one-way ANOVA followed by the Bonferroni post-test, two-sided Mann–Whitney test and two-sided Student’s; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Gray = vehicle; white = stimulation; orange=Foxo1;ADA blue=IL-6−/−. A detailed list of sample sizes and P value per graph is provided in the Supplemental Material. Source data are provided as a Source Data file.

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