Journal: Mediators of Inflammation

Article Title: CRP Stimulates GDF15 Expression in Endothelial Cells through p53

doi: 10.1155/2018/8278039

Figure Lengend Snippet: Women with hsCRP have high levels of GDF15. (a, b) Serum GDF15 levels were quantified by ELISA from HANDLS participants with either low- (<3 mg/L), mid- (>3–20 mg/L), or high hsCRP (>20 mg/L) levels ( n = 39/group). The ELISA assay was performed according to manufacturer's instructions and was repeated in 2 independent experiments. (c) RNA was isolated from PBMCs from HANDLS participants with either low- (<3 mg/L) or high hsCRP (>20 mg/L) levels ( n = 15/group). GDF15 mRNA was quantified by RT-qPCR and normalized to HPRT1 and UBC levels. The histograms represent the mean + SEM from three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 by Student's t -test.

Article Snippet: Human GDF15 Quantikine ELISA Kit (R&D Systems) was used according to the manufacturer's directions.

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Quantitative RT-PCR

Journal: Mediators of Inflammation

Article Title: CRP Stimulates GDF15 Expression in Endothelial Cells through p53

doi: 10.1155/2018/8278039

Figure Lengend Snippet: CRP upregulates GDF15 expression. (a) 18 h after CRP treatment with the indicated doses, GDF15 expression in HAECs was analyzed by immunoblotting with anti-GDF15 antibodies. β -Actin was used as a loading control. (b) After CRP treatment for the indicated time points, conditioned media was collected and GDF15 secreted levels were analyzed by ELISA. GDF15 levels were normalized to the 0 h time point for each experiment. The mean of three independent experiments is shown. (c and d) 18 h after CRP (25 μ g/mL) treatment, HAECs were lysed and levels of GDF15 mRNA or protein were quantified by RT-qPCR analysis (c) and western blot analysis (d). (e) HAECs were transfected with pCMV6-control or pCMV6-CRP plasmid for 48 h. Total RNA was isolated, and mRNA levels were quantified using RT-qPCR and normalized to GAPDH . (f) Total cell lysates from the indicated transfected HAECs were analyzed by Western blotting. β -Actin was used as a loading control. The histograms represent the mean + SEM from three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 by Student's t -test.

Article Snippet: Human GDF15 Quantikine ELISA Kit (R&D Systems) was used according to the manufacturer's directions.

Techniques: Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Transfection, Plasmid Preparation, Isolation

Journal: Mediators of Inflammation

Article Title: CRP Stimulates GDF15 Expression in Endothelial Cells through p53

doi: 10.1155/2018/8278039

Figure Lengend Snippet: CRP promotes GDF15 transcription. (a) Schematic of GDF15 promotor dual-luciferase constructs. Two p53 binding sites are indicated. (b) The indicated plasmids (1 μ g) were cotransfected with 0.1 μ g of TK-Renilla reporter plasmid in HeLa cells, and 24 h later, the cells were treated with CRP. After 18 h, the promoter activities were measured by luciferase activity. Transfection efficiency for luciferase activity was normalized to the Renilla luciferase activity. The results show the mean + SEM of three independent transfections. ∗∗ p < 0.01 by Student's t -test. (c) Schematic of p53 binding sites and primers used for ChIP assays in the GDF15 promoter. (d) ChIP assays were performed on HAECs transfected for 24 h and treated with or without CRP for 18 h. DNA immunoprecipitated by antibodies to p53 or immunoglobulin G IgG (control) was amplified by qPCR. Each qPCR reaction was performed in triplicate, and the histogram represents the average of three independent ChIP assays + SEM.

Article Snippet: Human GDF15 Quantikine ELISA Kit (R&D Systems) was used according to the manufacturer's directions.

Techniques: Luciferase, Construct, Binding Assay, Plasmid Preparation, Activity Assay, Transfection, Immunoprecipitation, Control, Amplification

Journal: Mediators of Inflammation

Article Title: CRP Stimulates GDF15 Expression in Endothelial Cells through p53

doi: 10.1155/2018/8278039

Figure Lengend Snippet: p53 knockdown inhibits CRP-induced GDF15 expression. HAECs were transfected with either Ctrl siRNA or p53 siRNA for 24 h and treated with or without CRP for 18 h. GDF15 mRNA levels were examined by RT-qPCR (a), and protein levels were analyzed by Western blot analysis (b). The histogram represents the mean + SEM from three independent experiments. ∗ p < 0.05 and ∗∗ p < 0.01 by Student's t -test.

Article Snippet: Human GDF15 Quantikine ELISA Kit (R&D Systems) was used according to the manufacturer's directions.

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

Journal: Nature Communications

Article Title: Transgenerational inheritance of diminished ovarian reserve triggered by prenatal propylparaben exposure in mice

doi: 10.1038/s41467-025-63440-z

Figure Lengend Snippet: a Representative ovarian images for TUNEL staining of F1–F3 offspring mice in Con and PrP groups at 12 w. b The number of TUNEL-positive cells per 1 mm 2 in ovarian sections of F1–F3 offspring mice in Con and PrP groups ( n = 5 biological replicates). Data are presented as mean ± SEM (unpaired two-tailed t -test). c The number of abnormal mitochondria in mice ovaries of F1–F3 generation ( n = 3 biological replicates, two non-overlapping microscope fields per ovary). Data are presented as mean ± SEM (unpaired two-tailed t -test). d Representative TEM images of mice ovaries in F3 generation. Yellow arrow: synapses of granulosa cells. M: mitochondria. The experiment was repeated three times. e Schematic illustration of the ovulation induction. f Representative oocyte images after ovulation induction. g The number of oocytes of F1 and F3 offspring mice in Con and PrP groups after ovulation induction (F1 n = 9; F3 Con n = 6, PrP n = 5 biological replicates). Data are presented as mean ± SEM (unpaired two-tailed t -test). h Representative IHC images showing immunoreactivity against BMP15 in F1, F2 and F3 offspring ovaries. i IHC scores based on IOD/Area of anti-BMP15 in F1, F2 and F3 offspring ovaries ( n = 5 biological replicates). Data are presented as mean ± SEM (unpaired two-tailed t -test). Source data are provided as a Source Data file. Figure 2e is created in BioRender. Wang, S. (2025) https://BioRender.com/bz8sq44 .

Article Snippet: The Antibodies were purchased from different companies: Cell Signaling Technology for ERK (Extracellular Signal-Regulated Kinase; 4695T), p-ERK (Phosphorylated ERK; 4370T), p38 (p38 Mitogen-Activated Protein Kinase; 8690T), p-p38 (Phosphorylated p38; 4511T), JNK (c-Jun N-terminal Kinase; 9252T), and p-JNK (Phosphorylated JNK; 4668T); Abclonal for β-ACTIN (AC038), active-Caspase3 (A19654), BCL-2 (A19693), BAX (A12009), SOD2 (A19576), BMP15 (A7321), CAT (A11777), HSD17B7 (A17157), and CTSK (A1782); Proteintech for FGF18 (60341-1-lg); Immunoway for RhoBTB1 (YT4084); Servicebio for HRP conjugated goat anti-rabbit IgG (GB23303), and Alexa Fluor® 488-conjugated goat anti-rabbit IgG (GB25303).

Techniques: TUNEL Assay, Staining, Two Tailed Test, Microscopy