Journal: AMB Express

Article Title: Treponema pallidum inhibits CD4+ T-cell proliferation through METAP2: insights from Mendelian randomization analysis

doi: 10.1186/s13568-025-01940-3

Figure Lengend Snippet: METAP2 as a potential key molecule in regulating CD4 + T-cells in NS. A Venn diagram showing the overlap between cis-proteins from rQTLs identified via mediation analysis and RNA-seq data from CD4 + T-cells of NS patients ( GSE103599 ). B The relationships between DelncRNAs and DEmiRNAs identified in GSE103599 and GSE156421 were determined via the starBase v3.0 database. The interactions between DEmiRNAs and METAP2 were predicted via TargetScan, miRcode, and MiRanda. On the basis of the predicted interactions among lncRNAs, miRNAs, and mRNAs, a ceRNA regulatory network was constructed and visualized via Cytoscape 3.10.2. C qRT‒PCR analysis was used to detect changes in the mRNA expression levels of LINC00501, LINC00334, METAP2, and PDCD5 in Jurkat T-cells treated with LTP, DTP, or PBS for 24 h. D Western blotting was performed to assess changes in the protein expression levels of METAP2 and PDCD5 in Jurkat T-cells treated with LTP, DTP, or PBS for 24 h. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant

Article Snippet: The membranes were blocked with 5% nonfat dry milk and incubated overnight at 4 °C with antibodies against METAP2 (#sc-365637, Santa Cruz Biotechnology), PDCD5 (#YN1785, Immunoway), and ACTB (#sc-47778, Santa Cruz Biotechnology).

Techniques: RNA Sequencing, Construct, Expressing, Western Blot

Journal: AMB Express

Article Title: Treponema pallidum inhibits CD4+ T-cell proliferation through METAP2: insights from Mendelian randomization analysis

doi: 10.1186/s13568-025-01940-3

Figure Lengend Snippet: T. pallidum inhibits CD4 + T-cell proliferation through METAP2. A-D qRT-PCR and Western blotting were used to measure METAP2 mRNA and protein expression levels in Jurkat T-cells 48 h after METAP2 knockdown or overexpression to assess the transfection efficacy. E , F Flow cytometry was used to assess Ki67 levels to evaluate the impact of METAP2 gene knockdown or overexpression on Jurkat T-cell proliferation. G Flow cytometry was used to determine Ki67 levels to assess the effects of METAP2 overexpression and T. pallidum treatment on Jurkat T-cell proliferation. ** p < 0.01; *** p < 0.001; ns, not significant

Article Snippet: The membranes were blocked with 5% nonfat dry milk and incubated overnight at 4 °C with antibodies against METAP2 (#sc-365637, Santa Cruz Biotechnology), PDCD5 (#YN1785, Immunoway), and ACTB (#sc-47778, Santa Cruz Biotechnology).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Knockdown, Over Expression, Transfection, Flow Cytometry

Journal: Genomics, Proteomics & Bioinformatics

Article Title: The Proteome Landscape of Human Placentas for Monochorionic Twins with Selective Intrauterine Growth Restriction

doi: 10.1016/j.gpb.2023.03.002

Figure Lengend Snippet: Impairment of angiogenesis in IUGR twin placentas A. IPA identified the “angiogenesis” annotation to be affected in IUGR twin placentas. * indicates that multiple identifiers in the dataset file map to a single gene in the Global Molecular Network. B. Immunostaining of CD34 in human placentas from IUGR twins and normal cotwins. CD34 is limited to endothelial cells. Scale bar, 50 µm. C. Placental MVD in IUGR twins and normal cotwins. D. Placental vascular area density in IUGR twins and normal cotwins. E. Immunohistochemical images of EFNB2, VIM, and METAP2 in placentas from IUGR twins and normal cotwins. Scale bar, 100 µm. F. – H. Immunostaining intensity of EFNB2 (F), VIM (G), and METAP2 (H) per tissue area. The data are represented by the fold decrease relative to the normal control mean. I. – K. qRT-PCR analysis showed that the mRNA levels of EFNB2 (I), VIM (J), and METAP2 (K) were significantly lower in the placentas of IUGR twins than in those of normal cotwins. All data are shown as mean ± SD with six samples per group. *, P < 0.05; **, P < 0.01; ***, P < 0.001. MVD, microvessel density; NC, negative control; SD, standard deviation.

Article Snippet: After deparaffinization, antigen retrieval, and endogenous peroxide blocking, the sections (5 μm in thickness) were incubated with primary anti-CD34 antibody (1:200; Catalog No. ZM-0046, ZSGB-BIO, Beijing, China), anti-EFNB2 antibody (1:50; Catalog No. HPA008999, Atlas Antibodies, Bromma, Sweden), anti-METAP2 antibody (1:3000; Catalog No. HPA019095, Atlas Antibodies), anti-VIM antibody (1:200; Catalog No. ZM-0260, ZSGB-BIO) and anti-MTDH antibody (1:150; Catalog No. ab124789, Abcam, Cambridge, UK).

Techniques: Immunostaining, Immunohistochemical staining, Control, Quantitative RT-PCR, Negative Control, Standard Deviation