oga  (Affinity Biosciences)

Journal: Molecular Medicine Reports

Article Title: miRNA-29a inhibits the proliferation of HUVECs by regulating the ITGB1/β-catenin/c-Myc pathway

doi: 10.3892/mmr.2026.13825

Figure Lengend Snippet: Inhibition of OGT downregulates the c-Myc pathway in HUVECs. HUVECs were transfected with 20 nM OGT siRNA1, siRNA2 or siRNA negative control for 6 h and cultured for another 48 h. The gene expressions of (A) OGT, (B) OGA and (C) c-Myc were detected using reverse transcription-quantitative PCR. (D) OGT, OGA and c-Myc protein expression levels were assessed using western blotting and (E) semi-quantified. (F) Cell viability was analyzed via Cell Counting Kit-8 assay. n=3; **P<0.01 compared with siRNA control group. HUVECs, human umbilical vein endothelial cells; OGT, O-linked N-acetylglucosamine transferase 110 kDa subunit; OGA, β-N-acetylglucosaminidase; siRNA, small interfering RNA.

Article Snippet: The membranes were blocked with 5% skimmed milk for 1 h at room-temperature and incubated with the following primary antibodies overnight at 4°C: ITGB1 (cat. no. 12594-1-AP; Proteintech Group, Inc.), T-cell factor 1 (TCF1; cat. no. 14464-1-AP; Proteintech Group, Inc.), EZH2 (cat. no. ab307646; Abcam), c-Myc (cat. no. 67447-1-Ig; Proteintech Group, Inc.), phosphorylated (p-)GSK3β (cat. no. AF2016; Affinity Biosciences), GSK3β (cat. no. AF5016; Affinity Biosciences), p-β-catenin (cat. no. DF2989; Affinity Biosciences), β-catenin (cat. no. 17565-1-AP; Proteintech Group, Inc.), β-actin (cat. no. 66009-1-Ig; Proteintech Group, Inc.), OGT (cat. no. ab177941; Abcam) and OGA (cat. no. DF8436; Affinity Biosciences).

Techniques: Inhibition, Transfection, Negative Control, Cell Culture, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Cell Counting, Control, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: miRNA-29a inhibits the proliferation of HUVECs by regulating the ITGB1/β-catenin/c-Myc pathway

doi: 10.3892/mmr.2026.13825

Figure Lengend Snippet: miR-29a downregulates the β-catenin/c-Myc pathway in HUVECs by targeting ITGB1. HUVECs were transfected with 20 µg/ml miR-29a mimics, miR-29a inhibitor or negative control for 6 h and cultured for another 48 h. (A) The protein expression levels of OGT, OGA, ITGB1, TCF1, c-Myc, p-GSK3β, GSK3β, p-β-catenin and β-catenin were assessed using western blotting and (B) semi-quantified. p-GSK3β and p-β-catenin were normalized to the levels of total GSK3β or β-catenin, respectively. (C) Cell viability was analyzed using a Cell Counting Kit-8 assay. The interaction between miR-29a and ITGB1 was validated using (D) TargetScan and (E) via dual-luciferase reporter assay (**P<0.01 compared with miR NC + ITGB1 WT group). (F) A schematic figure illustrating the interactions among miR-29a, ITGB1 and the c-Myc pathway. n=3; *P<0.05 and **P<0.01 compared with miRNA-NC group. miR/miRNA, microRNA; HUVECs, human umbilical vein endothelial cells; ITGB1, integrin β1; OGT, O-linked N-acetylglucosamine transferase 110 kDa subunit; OGA, β-N-acetylglucosaminidase; TCF1, T-cell factor 1; p- phosphorylated; hsa-miR, human microRNA; NC, negative control; WT, wild-type; Mut, mutant.

Article Snippet: The membranes were blocked with 5% skimmed milk for 1 h at room-temperature and incubated with the following primary antibodies overnight at 4°C: ITGB1 (cat. no. 12594-1-AP; Proteintech Group, Inc.), T-cell factor 1 (TCF1; cat. no. 14464-1-AP; Proteintech Group, Inc.), EZH2 (cat. no. ab307646; Abcam), c-Myc (cat. no. 67447-1-Ig; Proteintech Group, Inc.), phosphorylated (p-)GSK3β (cat. no. AF2016; Affinity Biosciences), GSK3β (cat. no. AF5016; Affinity Biosciences), p-β-catenin (cat. no. DF2989; Affinity Biosciences), β-catenin (cat. no. 17565-1-AP; Proteintech Group, Inc.), β-actin (cat. no. 66009-1-Ig; Proteintech Group, Inc.), OGT (cat. no. ab177941; Abcam) and OGA (cat. no. DF8436; Affinity Biosciences).

Techniques: Transfection, Negative Control, Cell Culture, Expressing, Western Blot, Cell Counting, Luciferase, Reporter Assay, Mutagenesis

Journal: The Journal of Prevention of Alzheimer's Disease

Article Title: Synaptic toxicity of OGA inhibitors and the failure of ceperognastat

doi: 10.1016/j.tjpad.2025.100456

Figure Lengend Snippet: OGA inhibitors alter synaptic plasticity. A , sample traces of whole-cell patch clamp recording of hippocampal CA1 pyramidal neurons during the first episode of paired-pulse stimulation ( A1 ) and aligned first response from every episode ( A2 ). Scale bar, 20 ms (horizontal) and 0.5pA (vertical). The color coding for DMSO (black), ASN90 ( i.e. , ASN, red), LY3372689 ( i.e. , LY, green), and MK8719 ( i.e. , MK, blue) is used in all plots. B , plots of EPSC ratio for the responses during the first episode ( B1 ) and the first responses of five episodes ( B2 ). C , LTP measured as relative EPSP slopes after the treatments of OGAi or DMSO. Left insets are sample traces pre- and post 100-Hz 1-s tetanus stimulation. Right inset is a violin plot of the average values during the last 30-s recording ( i.e. , gray box in the plot). Welch’s ANOVA test, W (DFn, DFd) = 10.61 (3.000, 17.50), p = 0.0003; Kolmogorov-Smimov test, p = 0.0063 (ASN vs. DMSO), 0.0336 (LY vs. DMSO), and 0.0063 (MK vs. DMSO). N = 3 mice and n = 3 slices ( i.e. , 9 recordings in total for every treatment). *, 0.05 > p ≥ 0.01; **, 0.01 > p ≥ 0.005; ***, 0.005 > p ≥ 0.001, ****, p < 0.001.

Article Snippet: As Biogen’s Phase I BIIB113 study [ ] highlighted, potential impacts on synaptic transmission should be considered in the design and interpretation of clinical OGA inhibitor programs.

Techniques: Patch Clamp

Journal: The Journal of Prevention of Alzheimer's Disease

Article Title: Synaptic toxicity of OGA inhibitors and the failure of ceperognastat

doi: 10.1016/j.tjpad.2025.100456

Figure Lengend Snippet: OGA inhibitors alter synaptic protein levels and Tau phosphorylation. A , sample images of fluorescence immunocytochemistry for PSD95, Tuj1, GFAP, and DAPI. Scale bar, 0.5 mm. Images on the left are zoom-in images of PSD95 immunofluorescence in the cell-body layer (indicated by the white box) for all four treatments. Scale bar, 50 μm B , sample images of fluorescence immunocytochemistry for Synaptophysin 1, Tau, pTau, and DAPI. Images on the left are zoom-in images of Synaptophysin 1 immunofluorescence in the cell-body layer (indicated by the white box) for all four treatments. C & D , violin plots of PSD95 ( C ) and Synaptophysin 1 ( D ) immunofluorescence intensities. E , violin plots of pTau vs. Tau fluorescence intensity (F) ratios from the same ROIs of Synaptophysin 1. Welch’s ANOVA test for PSD95, W (DFn, DFd) = 90.96 (3.000, 2168), p < 0.0001; for Synaptophysin 1 fluorescence intensity, W (DFn, DFd) = 153.30 (3.000, 2187), p < 0.0001; for pTau/Tau, W (DFn, DFd) = 128.40 (3.000, 1918), p < 0.0001. Kolmogorov-Smimov test for all three, p < 0.0001 (ASN vs. DMSO, LY vs. DMSO, and MK vs. DMSO). N = 9 slices from 3 mice (3 ea), n = 999 ROIs (111 randomly selected from every slice). *, 0.05 > p ≥ 0.01; **, 0.01 > p ≥ 0.005; ***, 0.005 > p ≥ 0.001, ****, p < 0.001.

Article Snippet: As Biogen’s Phase I BIIB113 study [ ] highlighted, potential impacts on synaptic transmission should be considered in the design and interpretation of clinical OGA inhibitor programs.

Techniques: Phospho-proteomics, Fluorescence, Immunocytochemistry, Immunofluorescence