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: medRxiv

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

doi: 10.1101/2022.08.29.22278892

Figure Lengend Snippet: A . IPA identified the “angiogenesis” annotation to be affected in IUGR twin placentas. 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 placental shares from IUGR twins and normal cotwins. Scale bar, 100 μm. F – H . EFNB2, VIM, and METAP2 immunostaining intensity per tissue area is represented by the fold-decrease relative to the normal control mean. I – K . qRT-PCR analysis showed that the EFNB2, VIM , and METAP2 transcriptional levels were significantly lower in placentas of IUGR twins compared with normal cotwins. All data are shown as mean ± SD with 6 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 (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 (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

Journal: British Journal of Cancer

Article Title: Disabled-2 downregulation promotes epithelial-to-mesenchymal transition

doi: 10.1038/sj.bjc.6605975

Figure Lengend Snippet: Human breast tumours exhibit decreased expression of the p96 and p67 forms of Dab2. Amplified cDNA prepared from 12 normal and 12 breast tumour samples arrayed in a multi-well PCR plate was obtained from the manufacturer (Origene). PCR analysis was carried out with primers flanking the splice site for generation of the p67 isoform of Dab2 and primers for human actin as control. Products were resolved on a 1% agarose gel and visualised by ethidium bromide staining. Amplification of the p96 isoform results in a product of 822 bp, whereas amplification of the p67 isoform yields a product of 167 bp. Images were obtained using the Kodak 1D programme following ultraviolet illumination. Histopathological data for individual BC samples is detailed in . Abbreviations: A=adenoid cystic; DCIS=ductal carcinoma in-situ , IDC=invasive ductal carcinoma; IT=invasive mixed tubular.

Article Snippet: Determination of p96 and p67 isoform expression levels in normal and breast tumour samples was performed utilising the Human Breast Cancer Rapid-Scan Panel (Origene) according to manufacturer's suggestions and performed in duplicate using the two individual plates provided.

Techniques: Expressing, Amplification, Agarose Gel Electrophoresis, Staining, In Situ

Journal: British Journal of Cancer

Article Title: Disabled-2 downregulation promotes epithelial-to-mesenchymal transition

doi: 10.1038/sj.bjc.6605975

Figure Lengend Snippet: Downregulation of Dab2 in HME5-cdk4 (K4) cells results in a constitutive EMT phenotype. ( A , B ) Cell lysates from unstimulated cells or cells treated with 2.5 ng ml −1 TGF β for 48 h were subjected to western blot analysis for Dab2, N-cadherin (Ncad) and vimentin expression as described in Materials and Methods. HSP90 expression by western blot analysis demonstrates equal protein loading ( n =3 independent experiments). Arrow indicates the position of the p96 Dab2 isoform; Asterisk indicates a non-specific band. ( C ) K4 Neg, K4 599 and K4 1059 cells were untreated or stimulated with TGF β (2.5 ng ml −1 ) for 48 h and subjected to western blot analysis for phospho-ERK1/2 (pERK1/2) and total ERK2 expression as described in Materials and Methods. ( D ) Elevated expression levels of TGF β isoforms exhibited by Dab2 knockdown cells. Transforming growth factor β 1, TGF β 2 and TGF β 3 mRNA expression was determined by qRT–PCR analysis in untreated K4 Neg and shDab2-expressing cells as described in Materials and Methods. Shown is mean±s.d. from assays performed in triplicate with expression in K4 Neg set to 1.0. ( n =3 independent experiments).

Article Snippet: Determination of p96 and p67 isoform expression levels in normal and breast tumour samples was performed utilising the Human Breast Cancer Rapid-Scan Panel (Origene) according to manufacturer's suggestions and performed in duplicate using the two individual plates provided.

Techniques: Western Blot, Expressing, Quantitative RT-PCR

Journal: British Journal of Cancer

Article Title: Disabled-2 downregulation promotes epithelial-to-mesenchymal transition

doi: 10.1038/sj.bjc.6605975

Figure Lengend Snippet: Loss of Dab2 facilitates EMT. In the basal state, cytosolic Grb2 can be found in association with Dab2 or Sos. Loss of Dab2 expression, commonly observed in breast cancer, is postulated to increase the proportion of Grb2 associated with Sos, resulting in Ras activation (RAS * ), stimulation of MAPK signalling and phosphorylation of ERK (pERK). Activation of ERK stimulates increased transcription of the TGF β isoforms, in particular the TGF β 2 isoform. Increased TGF β stimulation induces transient EMT, and also initiates establishment of an autocrine TGF β signalling loop leading to a stable EMT phenotype exemplified by increased expression of EMT markers. Inhibition of TGF β signalling with SB431542 or MAPK signalling with U0126 decreases TGF β production and causes partial reversion of the EMT phenotype.

Article Snippet: Determination of p96 and p67 isoform expression levels in normal and breast tumour samples was performed utilising the Human Breast Cancer Rapid-Scan Panel (Origene) according to manufacturer's suggestions and performed in duplicate using the two individual plates provided.

Techniques: Expressing, Activation Assay, Inhibition

Journal: Expert opinion on investigational drugs

Article Title: Obesity medications in development

doi: 10.1080/13543784.2020.1705277

Figure Lengend Snippet: Obesity Medications in the pipeline, their mechanism of action, stage of development, and sponsor

Article Snippet: [ 99 , 100 ] The MetAP2 inhibitor beloranib (ZGN-440, Zafgen), consistently produced clinically significant weight loss in patients with obesity, type 2 diabetes and Prader Willi syndrome, with 13% weight loss over 26 weeks in patients with diabetes.

Techniques: Medications