Fig. 3: PEAR1 associated with PP1 to suppress fibrotic factor induced fibroblast activation. | Nature Communications

Fig. 3: PEAR1 associated with PP1 to suppress fibrotic factor induced fibroblast activation.

From: PEAR1 regulates expansion of activated fibroblasts and deposition of extracellular matrix in pulmonary fibrosis

Fig. 3

Pdgfra+ cells were selected for bulk RNA-seq assay. a Overlapping pattern of the differentially expressed genes (DEGs) in three comparisons identified 155 core DEGs. b Principle component analysis (PCA) of replicated samples in each condition. Squares represent the centroid of corresponding sample group. c Top 10 GO biological process enriched with upregulated genes in Pear1−/− Bleo compared to WT Bleo. d Clustering of the leading-edge EMT genes in GSEA across 12 samples in four experiment conditions. Colored squares on bottom left represent experiment conditions. Colored bar on bottom right represents the scaled expression levels. e Leading-edge EMT genes in GSEA were predominantly expressed in cluster 7 (activated fibroblasts) cells of scRNA-seq data. Color bar on bottom left represents the scaled expression levels. Color dot on bottom left represents Pdgfra+ cell clusters in scRNA-seq data: 1-Col14a1 Matrix Fibroblasts, 2-Col14a1/Col13a1 Matrix Fibroblasts, 5-Col13a1 Matrix Fibroblasts, 7-Activated Fibroblasts, 8-Col14a1 Matrix Fibroblasts (Mmp3/Cyp1b1 high). f The phosphorylation levels of Smad2/3, AKT, ERK1/2, P38, and JNK1/2 were evaluated in WT and Pear1−/− fibroblasts stimulated by TGFβ, FGF, or PDGF for 30 min, respectively. GAPDH was used as a loading control. The experiments were repeated three times and the results were similar. g, h The binding of PEAR1 and PP1 were detected by immunoprecipitation in cultured human and mouse pulmonary fibroblasts. The experiments were repeated three times and the results were similar. i The phosphorylation levels of, AKT, P38, ERK1/2, and JNK1/2 were evaluated in fibroblasts incubated with 1 nM, 10 nM, or 100 nM PP1 inhibitor for 15 min, respectively. GAPDH was used as a loading control. The experiments were repeated three times and the results were similar. j The relative mRNA expression levels of ECM genes in cultured pulmonary fibroblasts incubated with PP1 inhibitor or DMSO as a negative control (n = 3 biologically independent samples in each group). For c, one-sided Fisher’s exact test was used. For j, two-tailed t test was used. Data are presented as mean ± SD. Source data are provided as a Source data file.

Back to article page