Journal: Cell Death Discovery

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

doi: 10.1038/s41420-024-02048-6

Figure Lengend Snippet: 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.

Article Snippet: CCL2 , Cell Signaling Technology, 39091 , 1:1000 , WB , R , AB_2799147.

Techniques: RNA Sequencing, Cell Culture, Control, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: Cell Death Discovery

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

doi: 10.1038/s41420-024-02048-6

Figure Lengend Snippet: A After 48 h incubation, Western blotting shows that IL-1β increased IL-1β, IL-6, TNF-α, MMP3, p16, p21, CCL2 and phospho-histone H2A.X in P3 ESC. B Time-course experiments reveal that IL-1β upregulated IL-1β, IL-6, TNF-α, MMP3, p16, p21, HMGB1, CCL2, CCL5 and phospho-histone H2A.X at 24, 48 and 72 h (lanes 2, 4 and 6). Co-incubation in the presence of the JNK inhibitor SP blocked IL-1β-mediated upregulation at all time points (lanes 3, 5 and 7). Incubation with SP alone (lane 8) suppressed many biomarkers below basal (Control, lane 1) levels. β-Actin levels were not affected. Molecular masses (kDa) are indicated by arrows at right. C Based on the time course experiments, the three key senescence-associated biomarkers indicated that the baseline levels of IL-1β and IL-6 were notably low and undetectable by the western blot method at all three time points. Meanwhile, MMP3 expression levels in the untreated control group remained consistently similar across the three observed time points. β-Actin levels were not affected. Molecular masses (kDa) are indicated by arrows at right.

Article Snippet: CCL2 , Cell Signaling Technology, 39091 , 1:1000 , WB , R , AB_2799147.

Techniques: Incubation, Western Blot, Control, Expressing

Journal: Cell Death Discovery

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

doi: 10.1038/s41420-024-02048-6

Figure Lengend Snippet: Antibody table.

Article Snippet: CCL2 , Cell Signaling Technology, 39091 , 1:1000 , WB , R , AB_2799147.

Techniques: Transfection

Journal: Nature Communications

Article Title: Transcriptional dynamics of murine motor neuron maturation in vivo and in vitro

doi: 10.1038/s41467-022-33022-4

Figure Lengend Snippet: a Principal component analysis on in vivo and in vitro datasets based on all distal ATAC-seq peaks. Each circle is the cumulative ATAC-seq data from two biological replicates. b ATAC-seq reads around a continuously expressed gene ( Chat ) and genes upregulated during maturation ( Spp1 and Thy1 ). c Heatmaps of ATAC-seq reads. Each panel spans ±1 kb from the center of peaks. Top: peaks upregulated both in vivo and in vitro around shared upregulated genes. Bottom: peaks upregulated only in vivo around genes upregulated only in vivo. d , e Motifs enriched in top ( d ) and bottom ( e ) genomic regions from ( c ). HOMER outputs show the de novo motif (top) and the best-matched known transcription factor motif (bottom) along with p value and prevalence. f Immunostaining of Nfia and Nfib in DIV28 cultures. g Immunostaining and scoring of Fos in DIV28 cultures in the absence and presence of TTX. All scale bars are 50 μm.

Article Snippet: The following primary antibodies were used: GFP (Chick, Thermo Fisher A10262, 1:3000), Chat (Goat, EMD Millipore AB144P 1:100), hb9 (Guinea pig 1:100 from Jessell Lab), NeuN (Rabbit, Millipore-Sigma ABN78, 1:1000), Spp1 (Mouse, R&D Systems AF808, 1:50; Goat, sc-21742, 1:300), Isl1/2 (Ms 4D5 DSHB optimized concentration by staining; Neuromics GT15051 1:5000); Nfia (Rb, Active Motif 39397, 1:1000), Nfib (Rb, Active Motif 39091, 1:1000), NFH (Chk, Neuromics CH22104, 1:2000), cFos (Rat, Synaptic Systems 226 017, 1:500), Nr3c1 (Rabbit, Invitrogen PA1-511A, 1:2000), Nr3c2 (DSHB clones 1D5 and 3F10, 1:33).

Techniques: In Vivo, In Vitro, Immunostaining