Research Paper Volume 15, Issue 22 pp 12998—13009

Figure 3. Gene expression profiling reveals an increase in genes associated with inflammation and RNA pull-down shows circFoxp1 interact with RACK1. (A) Heatmap and volcano plot showed the differentially expressed RNAs in paired samples of HSFs by RNA-seq analysis. Fold change > 1 or <−1.0, P value < 0.05. (B) GO annotations of the genes of upregulated expressed RNAs. The bar plot presented the enrichment scores (−loge [p value]) of the top 20 significantly enriched GO terms in biological processes, cellular components and molecular functions. (C) Bulb map of KEGG analysis for the genes of upregulated expressed RNAs. Rich factor represented the enrichment degree of differentially expressed genes. Y axis showed the name of enriched pathways. The area of each node represented the number of the enriched host genes of differentially expressed RNAs. The p-value was represented by a color scale. (D) Quantitative RT-PCR to quantify fibroblast migration marker (CD44), proliferation marker (Ki67) and inflammation cells marker (IL-6 and TNF-α) circFoxp1 overexpressed HSF cells. ^*P < 0.05, ^***P < 0.001. (E) ROS activity was determined by confocal fluorescence microscopy using CellROXTM Deep Orange Staining. Scale bar: 25 μm. ^****P < 0.0001. (F) RNA pulls down revealed several proteins interact with circFoxp1. (G) Western blot confirmed ROCK1 presented in RNA pulldown liquid. (H) ROCK1 reduced in the OE-circFoxp1 group as compared to OE-Ctr in HSFs.