Fig. 3: Identification of FoxO1-STAT3 signaling in bronchi, microvessels, and adjacent lung tissue from offspring at postnatal day 21 (P21).

A The heatmap from a transcriptomic analysis includes only genes with an absolute log-fold change (FC) > 2 (adjusted P value <0.05); standard diet (SD) and high-fat diet (HFD) represent the offspring of standard diet (SD)-fed dams (n = 2) and HFD-fed dams (n = 3), respectively. Z-Score is indicated in a color score. B Volcano plot analysis; genes with an absolute FC > 2 and adjusted P value <0.05 are indicated in red; genes with an FC > 2 but adjusted P value >0.05 are green (DEGs in SD compared to HFD). Labels in the volcano plot indicate cell cycle and FoxO1 target genes. P values are calculated using Wald test and P-adjusted values using the FDR/Benjamini–Hochberg approach. C, D Venn diagrams depicting the putative binding sites for the transcription factors Foxo1, STAT3, Jun, Nfe2l2, and Pparγ in the promoters of the top 100 upregulated and downregulated genes with adjusted P values <0.01. E Gene expression of Stat3, Socs3, and Jun in bronchial smooth muscle cells (bSMCs) isolated from offspring of SD- and HFD-fed dams at P21 (n = 4/group); Gapdh served as the housekeeping gene. F immunoblot for c-Jun and total STAT3 in total lung homogenates from HFD and SD at P21 (n = 6/group); a densitometric summary of the STAT3 and c-Jun data relative to β-actin is displayed next to the respective immunoblot. G A functional enrichment analysis to identify the upregulated biological processes (SD vs HFD) associated with differentially expressed genes (DEGs) with adjusted P values <0.05. A KEGG pathway analysis was used to identify the top ten upregulated pathways in the bronchi (SD vs HFD), microvessels, and adjacent lung tissue at P21. E, F Data are shown as mean ± standard error of the mean. The statistical analysis was conducted using the two-sided Mann–Whitney test and two-sided Student’s t test; *P < 0.05; ***P < 0.001. Gray = standard diet; white = high-fat diet. 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.