Fig. 5
From: TGF-β1-triggered maladaptive bone marrow endothelium impedes hematopoietic recovery

Temporal dissection of TGF-β1-regulated phosphorylation events. a Workflow of the phosphoproteome analysis. b Schematic overview of the time points used for TGF-β1 treatment (10 ng/ml). c Heatmap illustrating the outcomes of hierarchical clustering analysis based on z-scored intensities of phosphorylation (ANOVA test, P < 0.05). The phosphosites were clustered into seven categories according to their intensities. Seven clusters are also shown in the profile plot, with the numbers of included phosphosites indicated. d The bar graphs show the GOCC enriched in cluster 2, cluster 6 and cluster 7 of phosphoproteins. The analysis was performed using Perseus. e The similarity of the clustered pathways induced by TGF-β1. The profiles are color-coded according to their similarity from the respective cluster. The red scale represents the similarity score calculated via the WikiPathways analysis from the Cytoscape website, whereas gray represents missing values. f Volcano plots showing the indicated upregulated (red circles) and downregulated (blue circles) phosphosites after 12 h of TGF-β1 treatment. g HUVECs were treated for the indicated time periods with 10 ng/mL TGF-β1. The cell lysates were subjected to immunoblotting with anti-phospho-p38α (p-p38), anti-Fibronectin-1 (FN1) and anti-β-actin antibodies. The western blot analyses were performed in triplicate and representative images are shown