Fig. 1: RNA sequencing (RNAseq) and ELISA assays were used to confirm the senescence induced by IL-1β. | Cell Death Discovery

Fig. 1: RNA sequencing (RNAseq) and ELISA assays were used to confirm the senescence induced by IL-1β.

From: Interleukin-1β induces and accelerates human endometrial stromal cell senescence and impairs decidualization via the c-Jun N-terminal kinase pathway

Fig. 1

A The heatmap of RNA-Seq. P3 ESC from two subjects (S1, S2) were cultured without (-) or with (+) 0.1 nM IL-1β for 24 h and subjected to RNAseq for transcript profiling. The top 10 mRNAs upregulated by IL-1β are illustrated in the heat map. Red color indicates upregulated and blue color downregulated genes (fold changes shown in inset). B Volcano Plot RNA-Seq. A volcano plot demonstrates strong upregulation of SASP mRNA transcripts in P3 ESC from subjects S1 and S2 24 h after IL-1β treatment. IL-1β itself, IL-6, IL-8, CCL2, CCL5 and TNF-α mRNA were all increased over control and to a lesser extent, MMP3. C ELISA assay. ELISA of five SASP biomarker proteins in P3 ESC treated supernatants follow a time course after stimulation with IL-1β for up to 72 h. At the 72-h mark, ANOVA analysis revealed a significant overall difference (p < 0.001). The Scheffé Test results indicated significant differences between MMP3 and IL-8, IL-6, RANTES, and TNFα (p < 0.001 for all); between IL-8 and IL-6, RANTES, and TNFα (p > 0.05, p < 0.001, p < 0.001, respectively); and between IL-6 and RANTES, and TNFα (p < 0.001 for both), while for RANTES versus TNFα, there was no significant differences (p > 0.05). Each experiment was repeated three times. Error bars indicate SD of triplicate determinations. D ELISA assay. Data graphed with an expanded ordinate represent CCL5 and TNF-α, to adjust for relatively low absolute cytokine concentrations. Error bars indicate SD of triplicate determinations for CCL5. Other ELISA assays were done with duplicate samples.

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