eno3  (Sino Biological)

Journal: Cell Reports Medicine

Article Title: Non-orthosteric inhibition of enolase 1 impedes growth of triple-negative breast cancer

doi: 10.1016/j.xcrm.2025.102451

Figure Lengend Snippet: SU212 targets ENO1 (A) Chemical structure of podophyllotoxin (parental compound) and SU212. (B–D) SU212 binds to ENO1 and ENO3. Target identification using CETSA: Differential profiling of SU212 on the thermal proteome profile of MDA-MB-231 cells. Cells were treated with DMSO or SU212 (0.5 μM) for 1.5 h and then lysed, and an equal quantity of soluble protein was labeled with a tandem mass tag, followed by liquid chromatography-tandem mass spectrometry analysis. (B) Heatmap representation of the thermal stability of 1,074 soluble proteins in cancer cells treated with vehicle-DMSO (left) and SU212 (right). (C) A scatterplot of melting temperature (T m ) calculated after SU212 and vehicle treatment. Proteins that passed the significant value thresholds ( p < 0.01, R2 > 0.8) and identification criteria are highlighted in orange. (D) Melting curves for ENO1/ENO3 with and without SU212 treatment depict the change in T m . (E) Representative sensorgrams for ENO1/3-SU212 interaction. His-tagged ENO1 and ENO3 proteins were immobilized on a Ni-NTA sensor, and SU212 (10 μM) was tested for physical interaction using BLI. (F) SU212 physically interacts with ENO1 protein. The BLI sensorgrams were obtained using His tag-ENO1-loaded Octet NTA biosensors and SU212 (1, 5, 10 μM). (G) SU212 treatment inhibits ENO1 expression. Immunoblotting: TNBC cells were treated with vehicle only (DMSO) and SU212 (0.1, 0.25, 0.5 μM) for 6 h. SDS-PAGE and western blot analysis were performed for the ENO1 and ENO3 proteins. Membranes were stripped and re-probed with anti-beta-actin antibody to ensure equal protein loading. (H) SU212 treatment leads to degradation of the ENO1 protein. Immunoblotting: MDA-MB-231 cells were treated with combinations of SU212, CHX, MG132, 3MA, and NH4Cl, as depicted in the figure, for 6 h. SDS-PAGE and western blot analysis were performed for the ENO1 protein. Membranes were stripped and re-probed with anti-GAPDH antibody to ensure equal protein loading. (I) SU212 treatment induces apoptotic cell death in MDA-MB-231 cells. Data are shown as the mean ± SD ( n = 5). Numbers indicate a p value compared with the vehicle control and analyzed using two-way ANOVA. (J) Enolase enzyme activity assay. PC, positive control. Data are shown as the mean ± SD ( n = 3). Numbers indicate a p value that is different compared with vehicle control, analyzed using Student’s t test. ns, not significant.

Article Snippet: During the loading step, recombinant His Tag-Enolases were immobilized on NTA Biosensors using a kinetic buffer (1X PBS) with a final reagent volume of 50 μL, which contained 50 μg/mL of ENO1 (#11554-H07E−100, Sino Biological) and ENO3 (#14270-H07E, Sino Biological), all placed in a black 384-well microplate.

Techniques: Drug discovery, Labeling, Liquid Chromatography, Mass Spectrometry, Expressing, Western Blot, SDS Page, Control, Enzyme Activity Assay, Positive Control

Journal: Frontiers in Immunology

Article Title: Development of a diagnostic model for MASLD and identification of daidzein as the potential drug using bioinformatics analysis and experiments

doi: 10.3389/fimmu.2025.1698740

Figure Lengend Snippet: Identification of potential drugs for MASLD. (A) The bar plot displays the top 30 candidate drugs. (B) Molecular docking analysis of daidzein and ENO3 . (C) Chemical structure of daidzein. (D) CCK8 experiment. (E) Oil Red O and Bodipy 493/503 staining of primary hepatocytes. (F, G) Quantitative analysis by calculating the area of lipid droplets within cells. (H, I) Determination of (H) intracellular TG and (I) TC content. The experiment was repeated three times. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: ENO3 , Rabbit , Proteintech; 55234-1-AP.

Techniques: Staining

Journal: Frontiers in Immunology

Article Title: Development of a diagnostic model for MASLD and identification of daidzein as the potential drug using bioinformatics analysis and experiments

doi: 10.3389/fimmu.2025.1698740

Figure Lengend Snippet: Daidzein alleviates MASLD through the ENO3/PPAR signaling pathway. (A) Daidzein reduced the protein expression of ENO3 in primary hepatocytes. (B) GSEA analysis shows the top 10 pathways with high ENO3 expression. (C) Daidzein affects the expression of 3 proteins, PPARα, PPARγ, and PPARD, in primary hepatocytes. (D) Daidzein affects the protein expression of downstream proteins (SCD1,FASN, CD36 and CPT1A) in the PPAR pathway. (E, F) Perform protein quantification analysis of ENO3, PPAR, PPARγ PPARD, CPT-1A, CD36, FASN and SCD1. The experiment was repeated three times. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: ENO3 , Rabbit , Proteintech; 55234-1-AP.

Techniques: Expressing