Journal: Open Life Sciences

Article Title: TRIM28-mediated p53 ubiquitination inhibits trophoblast apoptosis to support normal placental development

doi: 10.1515/biol-2025-1236

Figure Lengend Snippet: Expression of TRIM28 and related indicators in placentas of L-NAME-induced EOPE mice. (A) Measurement of systolic blood pressure in mice using the tail-cuff method; (B) determination of urinary protein levels using the BCA assay; (C) HE staining observation of placental tissue structure, the scale bar represents 400 μm; (D) IHC detection of cleaved caspase-3, Bax, and Bcl-2 protein expression, the scale bar represents 100 μm; (E) measurement of ROS levels in placental tissue, the scale bar represents 200 μm; (F) analysis of TRIM28 mRNA expression in placenta by RT-qPCR; (G) IHC evaluation of TRIM28 protein expression in placental tissue, the scale bar represents 100 μm. N = 6. **** p < 0.0001.

Article Snippet: The lysates were incubated overnight at 4 °C with anti-TRIM28 antibody (3 μg, A2245, Abclonal) or control rabbit immunoglobulin G (1 μg, 30000-0-AP, Proteintech, Wuhan, China) pre-coupled to Protein A/G PLUS-Agarose beads (20423, Thermo Fisher Scientific).

Techniques: Expressing, BIA-KA, Staining, Quantitative RT-PCR

Journal: Open Life Sciences

Article Title: TRIM28-mediated p53 ubiquitination inhibits trophoblast apoptosis to support normal placental development

doi: 10.1515/biol-2025-1236

Figure Lengend Snippet: Alleviation of L-NAME-induced placental injury by TRIM28 overexpression. (A) RT-qPCR detection of TRIM28 mRNA expression in placental tissue; (B) IHC detection of TRIM28 protein expression in placenta, the scale bar represents 100 μm; (C) measurement of systolic blood pressure in pregnant mice; (D) urinary protein detection to evaluate renal impairment; (E) HE staining for histological observation of placental structure, the scale bar represents 400 μm; (F) IHC detection of cleaved caspase-3, Bax, and Bcl-2 expression to assess apoptosis, the scale bar represents 100 μm; (G) measurement of ROS levels in placental tissue to evaluate oxidative stress, the scale bar represents 200 μm. N = 6. **** p < 0.0001.

Article Snippet: The lysates were incubated overnight at 4 °C with anti-TRIM28 antibody (3 μg, A2245, Abclonal) or control rabbit immunoglobulin G (1 μg, 30000-0-AP, Proteintech, Wuhan, China) pre-coupled to Protein A/G PLUS-Agarose beads (20423, Thermo Fisher Scientific).

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Staining

Journal: Open Life Sciences

Article Title: TRIM28-mediated p53 ubiquitination inhibits trophoblast apoptosis to support normal placental development

doi: 10.1515/biol-2025-1236

Figure Lengend Snippet: Reduction of H 2 O 2 -induced apoptosis in HTR-8/SVneo cells by TRIM28 overexpression. (A) RT-qPCR detection of TRIM28 mRNA expression; (B) WB detection of TRIM28, cleaved caspase-3, Bax, and Bcl-2 protein expression; (C) CCK-8 assay for evaluation of cell viability; (D) TUNEL staining for detection of apoptotic cells, the scale bar represents 200 μm; (E) ROS level detection, the scale bar represents 200 μm. N = 3. *** p < 0.001, **** p < 0.0001.

Article Snippet: The lysates were incubated overnight at 4 °C with anti-TRIM28 antibody (3 μg, A2245, Abclonal) or control rabbit immunoglobulin G (1 μg, 30000-0-AP, Proteintech, Wuhan, China) pre-coupled to Protein A/G PLUS-Agarose beads (20423, Thermo Fisher Scientific).

Techniques: Over Expression, Quantitative RT-PCR, Expressing, CCK-8 Assay, TUNEL Assay, Staining

Journal: Open Life Sciences

Article Title: TRIM28-mediated p53 ubiquitination inhibits trophoblast apoptosis to support normal placental development

doi: 10.1515/biol-2025-1236

Figure Lengend Snippet: Upregulation of p53 in placentas of L-NAME-treated mice and H 2 O 2 -stimulated HTR-8/SVneo cells. (A) Prediction of TRIM28 downstream targets using the UbiBrowser database; (B) RT-qPCR detection of p53 mRNA expression in mouse placental tissue; (C) IHC detection of p53 protein expression in mouse placental tissue, the scale bar represents 100 μm; (D) RT-qPCR detection of p53 mRNA expression in H 2 O 2 -treated HTR-8/SVneo cells; (E) WB detection of p53 protein expression in H 2 O 2 -treated HTR-8/SVneo cells. N = 6 (B–C), N = 3 (D–E). *** p < 0.001, **** p < 0.0001.

Article Snippet: The lysates were incubated overnight at 4 °C with anti-TRIM28 antibody (3 μg, A2245, Abclonal) or control rabbit immunoglobulin G (1 μg, 30000-0-AP, Proteintech, Wuhan, China) pre-coupled to Protein A/G PLUS-Agarose beads (20423, Thermo Fisher Scientific).

Techniques: Quantitative RT-PCR, Expressing

Journal: Open Life Sciences

Article Title: TRIM28-mediated p53 ubiquitination inhibits trophoblast apoptosis to support normal placental development

doi: 10.1515/biol-2025-1236

Figure Lengend Snippet: Ubiquitination regulation of p53 by TRIM28. (A) WB analysis of p53 protein expression in oe-TRIM28 and oe-NC groups; (B) Co-IP detection of TRIM28–p53 interaction; (C) Co-IP detection of p53–Ub binding and assessment of TRIM28’s effect on p53 ubiquitination; (D) evaluation of p53 protein stability following CHX treatment. N = 3. *** p < 0.001, **** p < 0.0001.

Article Snippet: The lysates were incubated overnight at 4 °C with anti-TRIM28 antibody (3 μg, A2245, Abclonal) or control rabbit immunoglobulin G (1 μg, 30000-0-AP, Proteintech, Wuhan, China) pre-coupled to Protein A/G PLUS-Agarose beads (20423, Thermo Fisher Scientific).

Techniques: Ubiquitin Proteomics, Expressing, Co-Immunoprecipitation Assay, Binding Assay