Journal: Aging Cell

Article Title: The Proinflammatory Secretome of Senescent Cells Can Be Controlled by a HIF2A ‐Dependent Upregulation and a FURIN ‐Dependent Cleavage of the ANGPTL4 Secreted Factor

doi: 10.1111/acel.70307

Figure Lengend Snippet: Cleavage of ANGPTL4 by FURIN is required for its ability to promote the proinflammatory SASP. (A‐C) MRC5/RAF:ER cells were transfected with control (siCTRL) or FURIN (siFURIN) siRNA and treated (+) or not (−) with 4‐OHT to induce senescence (OIS). (A) RT‐qPCR analysis against the indicated genes 3 days after 4‐OHT treatment. Mean ± SEM of n = 3 independent experiments. Paired one‐way ANOVA test results are shown. (B) Western blot analysis of FURIN, ANGPTL4: Full length in whole cell extracts (ANGPTL4) and secreted in the supernatant (cANGPTL4) and IL6. Representative picture of n = 3 independent experiments. (C) RT‐qPCR analysis of the indicated SASP encoding genes. Mean ± SEM of n = 3 independent experiments. Paired one‐way ANOVA test values are shown. (D) Relative mRNA expression of FURIN , ANGPTL4 and proinflammatory SASP encoding genes measured by RT‐qPCR in MRC5 cells overexpressing or not ANGPTL4 (ANGPTL4 OE) and transfected with control (siCTRL) or FURIN (siFURIN) siRNA. Mean ± SEM of n = 4 independent experiments. Paired one‐way ANOVA test results are shown.

Article Snippet: The membranes were then blocked for 1 h with 5% milk in TBS‐T 0.05% and subsequently incubated overnight at 4°C with primary antibodies in TBS‐T with 5% milk: α‐ANGPTL4 (sc‐373761, Santa Cruz, 1/250), α‐IL6 (sc‐28343, Santa Cruz, 1/500), α‐FURIN (18413‐1‐AP, Proteintech, 1/1000), α‐TUBULIN (T6199‐100, Sigma, 1/5000). α‐HIF1A (610958, BD Biosciences, 1/1000), HIF2A (NB100‐132, Novus Biologicals, 1/1000), INHBA (GTX108405, GeneTex 1/1000).

Techniques: Transfection, Control, Quantitative RT-PCR, Western Blot, Expressing

Journal: Communications Biology

Article Title: CCAAT/enhancer binding protein beta (C/EBPβ) regulates the formation of the ovarian reserve

doi: 10.1038/s42003-025-08798-y

Figure Lengend Snippet: A The distribution of genomic peaks in the Ctrl and siEp300 groups. B Column graph of DARs between Ctrl and siEp300 groups. C Genomic distribution of downregulated DARs in the siEp300 vs . Ctrl groups. D Venn diagram of the gene set with down-regulated DARs in the promoter region, the DEG set up-regulated at PD0, and the DEG set up-regulated at PD3. E Metascape enrichment analysis of candidate gene sets (Gene set acquisition has been described in the text). F Dot plot shows the expression of NTRK signaling-related genes in metascape enrichment analysis in E16.5, PD0, and PD3 oocytes. G The normalized reads of NTRK core-related genes Furin , Pcsk6 , and Rap1a shown by ATAC-seq. H Analysis of mRNA expression levels of Furin , Pcsk6 , and Rap1a in the Ctrl and siEp300 groups. I Ranking of TFs binding to the Furin promoter region obtained using The Signaling Pathways Project by binding score. J Detection of the binding activity of C/EBPβ to the Furin promoter region in the Ctrl and siEp300 groups using CUT&RUN-qPCR. K Detection of the enrichment level of Furin promoter fragment bound to p300 using CUT&RUN-qPCR. L Representative images (left) and expression level analysis (right) of FURIN, DDX4, LHX8, and SOHLH1 proteins in the Ctrl and siFurin groups. Violin plots showing the percentage of oocytes in follicles ( M ) and the total number of oocytes ( N ) in each ovarian section of each group. Data are means ± SEM. N ≥ 2 per group for the biological repeats of ATAC-seq; n = 5 to 6 for the biological repeats of CUT&RUN-qPCR, qPCR, WB, and oocyte count experiments, as shown on graphs.

Article Snippet: The membrane was blocked and incubated with the primary antibodies: DDX4 (ab13840, Abcam), C/EBPβ (ab32358, Abcam), LHX8 (ab137036, Abcam), SOHLH1 (orb158448, Biorbyt), NOBOX (NBP2-46193, Novus), GAPDH (AF7021, Affinity), p300 (ab275378, Abcam), FURIN (R382231, Zen-bioscience), p-C/EBPβ Thr188 (3084, Cell signaling technology), Lamin B1 (250010, Zen-bioscience) overnight at 4 °C.

Techniques: Expressing, Binding Assay, Protein-Protein interactions, Activity Assay

Journal: Communications Biology

Article Title: CCAAT/enhancer binding protein beta (C/EBPβ) regulates the formation of the ovarian reserve

doi: 10.1038/s42003-025-08798-y

Figure Lengend Snippet: A Schematic diagram of the mouse model of p300 inhibition and neurotrophin receptor activation. This figure was originally created by the authors using Adobe Illustrator. B Analysis of Furin mRNA levels in ovarian tissues of newborn mice treated with different concentrations of curcumin. C Analysis of Fshr mRNA levels in ovarian tissues of newborn mice treated with different concentrations of B-355252. D NGF levels in the serum of postpartum female mice in the Ctrl, Curcumin, Curcumin+B-355252, and B-355252 groups. E Representative images (left) and expression level analysis (right) of DDX4, LHX8, and SOHLH1 proteins in the Ctrl, Curcumin, Curcumin+B-355252, and B-355252 groups. F IF staining of DDX4 in the Ctrl, Curcumin, Curcumin+B-355252, and B-355252 groups. DDX4 (yellow) indicates oocytes. Hoechst (blue) indicates nuclei (Nuc). G , H Violin plots showing the percentage of oocytes in follicles ( G ) and the total number of oocytes ( H ) in each ovarian section of each group. Data are means ± SEM. N = 4 to 6 indicated the biological repeats, as shown on graphs.

Article Snippet: The membrane was blocked and incubated with the primary antibodies: DDX4 (ab13840, Abcam), C/EBPβ (ab32358, Abcam), LHX8 (ab137036, Abcam), SOHLH1 (orb158448, Biorbyt), NOBOX (NBP2-46193, Novus), GAPDH (AF7021, Affinity), p300 (ab275378, Abcam), FURIN (R382231, Zen-bioscience), p-C/EBPβ Thr188 (3084, Cell signaling technology), Lamin B1 (250010, Zen-bioscience) overnight at 4 °C.

Techniques: Inhibition, Activation Assay, Expressing, Staining

Journal: Communications Biology

Article Title: CCAAT/enhancer binding protein beta (C/EBPβ) regulates the formation of the ovarian reserve

doi: 10.1038/s42003-025-08798-y

Figure Lengend Snippet: A Representative images (left) and expression level analysis (right) of p-C/EBPβ T188 protein in the E16.5, PD0, PD3 ovaries. B Schematic diagram of the JNK inhibition (JNKi) model construction during in vitro culture of newborn ovarian. This figure was originally created by the authors using Adobe Illustrator. C Representative images of p-C/EBPβ T188 , C/EBPβ, p300, and FURIN proteins in the Ctrl and JNKi groups. Expression level analysis of p-C/EBPβ T188 ( D ), C/EBPβ ( E ), p300 ( F ), and FURIN ( G ) proteins in the Ctrl and JNKi groups. H IF staining of DDX4 in the Ctrl and JNKi groups. DDX4 (yellow) indicates oocytes. Hoechst (blue) indicates nuclei (Nuc). White arrow indicates the direction from the medulla (m) to the cortex (c). Violin plots showing the percentage of oocytes in follicles ( I ) and the total number of oocytes ( J ) in each ovarian section of each group. K Representative images (left) and line scan analysis (right) of the cellular localization of C/EBPβ in the Ctrl and JNKi groups. White dashed lines indicate the approximate outlines of the nests; white arrow indicates the direction from the medulla (m) to the cortex (c); yellow arrow indicates the direction of the line scan. L Fluorescence intensity analysis of the nuclei and cytoplasm of germ cells in the nests and follicles in the Ctrl and JNKi groups. M Proportion of oocytes with C/EBPβ N/C ratio > 1 in the total number of oocytes in the same ovarian section. Data are means ± SEM. N = 4–9 for the biological repeats of WB and oocyte count experiments, as shown on graphs; n = 6 for the biological repeats of cellular localization and fluorescence intensity analysis experiment, a dot in ( L ) represents a germ cell.

Article Snippet: The membrane was blocked and incubated with the primary antibodies: DDX4 (ab13840, Abcam), C/EBPβ (ab32358, Abcam), LHX8 (ab137036, Abcam), SOHLH1 (orb158448, Biorbyt), NOBOX (NBP2-46193, Novus), GAPDH (AF7021, Affinity), p300 (ab275378, Abcam), FURIN (R382231, Zen-bioscience), p-C/EBPβ Thr188 (3084, Cell signaling technology), Lamin B1 (250010, Zen-bioscience) overnight at 4 °C.

Techniques: Expressing, Inhibition, In Vitro, Staining, Fluorescence