dynamin related protein 1 drp1 inhibitor mdivi 1  (MedChemExpress)

Journal: Military Medical Research

Article Title: USP18 exacerbates myocardial I/R injury by inhibiting Parkin mitophagy through the deubiquitinase PTEN-L

doi: 10.1016/j.mmr.2026.100004

Figure Lengend Snippet: USP18 aggravates cardiac I/R injury through regulation of mitochondria and inhibition of mitophagy. a Electron microscopy image showing mitophagy in USP18-cKO mouse hearts ( n= 5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. b Protein levels of PINK1, Parkin, ubiquitinated proteins (Ub), P62, and LC3II in mitochondria from heart tissue in USP18-cKO and WT mice 24 h after I/R injury ( n =4). c Electron microscopy image showing mitophagy in USP18-overexpres (OV) mouse hearts ( n =5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. d Protein levels of PINK1, Parkin, Ub, P62, and LC3II proteins in mitochondria from the heart tissue of USP18-OV mice 24 h after I/R injury ( n =4). Color shift in mitophagy dye (red) and lysosomal dye (green) in NRVMs showing mitophagy in NRVMs with USP18 siRNA transfection ( e ) or Ad-USP18 infection ( f ) and the quantitative mitophagy index in each group ( n= 5). Scale bar=9 μm. Protein levels of PINK1, Parkin, Ub, P62, and LC3II in mitochondria from NRVMs transfected with USP18 siRNA ( g ) or infected with Ad-USP18 ( h ). ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001 ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WT. Wild-type; KO. Knockout; P62. Sequestosome 1; LC3. Microtubule-associated protein 1 light chain 3; VDAC. Voltage-dependent anion channel.

Article Snippet: To block mitophagy, the selective dynamin-related protein 1 (Drp1) inhibitor Mdivi-1 was used (50 μmol/L, MedChemExpress, USA).

Techniques: Inhibition, Electron Microscopy, Transfection, Infection, Ubiquitin Proteomics, Knock-Out

Journal: Pharmaceutical Science Advances

Article Title: Unleashing the power of DNA-encoded libraries for challenging targets in drug discovery

doi: 10.1016/j.pscia.2026.100115

Figure Lengend Snippet: Ligands targeting the GTPase KRAS G12C .

Article Snippet: In February 2025, Amgen reported a KRAS G12C covalent inhibitor.

Techniques:

Journal: MedComm

Article Title: Hypothalamus Participates in Peripheral Sympathetic Hyperactivity After Acute Brain Injury by Regulating Trhr M 6 A Modification

doi: 10.1002/mco2.71036

Figure Lengend Snippet: m 6 A modification by FTO participates in sympathetic hyperactivity after ABI. (A–C) Primary neurons were transfected with lentivirus encoding fast responsive Ca 2+ fluorescent probe jGCaMP7f in advance and then allocated to the sh‐Ctrl group, the sh‐FTO group, and the sh‐FTO+sh‐YTHDF2 group. The Basal Ca 2+ picture showed that no obvious change was observed in the sh‐Ctrl group, the sh‐FTO group, and the sh‐FTO+sh‐YTHDF2 group (A). Dynamic Ca 2+ detection revealed neural activation among the sh‐Ctrl group, the sh‐FTO group, and the sh‐FTO+sh‐YTHDF2 group (B and C). (D) Illustration of experimental process in vitro. (E–G) Representative pictures and analysis of immunoblot showed that neuron‐activation‐related proteins c‐FOS and p‐ERK were elevated in the TRH‐treated group compared to the control group, while decreased in the sh‐FTO+TRH group compared to the sh‐Ctrl+TRH group. (H) Illustration of experimental process in vivo. (I–K) Representative images and analysis of immunofluorescence co‐staining c‐FOS, Trhr, and NeuN showed that FB23 decreased the activation of neurons and the elevation of Trhr in the hypothalamus induced by TBI. (L–N) Sympathetic tone, parasympathetic tone, and stress index were obtained by analyzing HRV from mice in the sham group, the TBI group, and the TBI+FB23 group. (O) The concentration of plasma thyroid‐stimulating hormone (TSH) from the sham group and the TBI groups was detected by ELISA. (P–R) The concentrations of plasma TSH, norepinephrine, and ACTH from the sham group, the TBI group, and the TBI+FB23 group were detected by ELISA. n = 6. Data are presented as mean ± SEM and were analyzed by Student's t ‐test. ns represents no significance. * represents p < 0.05, ** represents p < 0.01, and *** represents p < 0.001. Scale bars: 25 µm.

Article Snippet: The selective FTO inhibitor FB23 (HY‐137187) and the thyrotropin‐releasing hormone (TRH, HY‐P0002) were purchased from MedChemExpress Co. (New Jersey, USA).

Techniques: Modification, Transfection, Activation Assay, In Vitro, Analysis, Western Blot, Control, In Vivo, Immunofluorescence, Staining, Concentration Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Journal of Animal Science and Biotechnology

Article Title: ZBED6 deletion activates FNDC1 -mediated PI3K/AKT and MAPK/ERK1/2 signaling pathways to promote myogenesis in pigs

doi: 10.1186/s40104-026-01504-w

Figure Lengend Snippet: ZBED6 deletion activates FNDC1 -mediated PI3K/AKT and MAPK/ERK1/2 signaling pathways. A RNA-seq data from ZBED6-deficient pigs, ZBED6 knockout myoblasts and myotubes, and FNDC1 knockdown C2C12 cells were comprehensively analyzed to screen downstream signaling pathways. B Protein expression levels and statistical analysis of p-AKT, AKT, p-mTOR and mTOR in WT and ZBED6 −/− pigs, n = 3 pigs/genotype. C and D Protein expression levels and statistical analysis, n = 3 pigs/genotype. E and F Protein expression levels and statistical analysis of p-mTOR, mTOR, p-AKT, AKT, p-ERK1/2 and ERK1/2, n = 3 independent cell-culture experiments. Data are expressed as mean ± SEM. In (B), and (D), the P values were determined by two-tailed Student's t -tests. In (F), the P values were determined by two-way ANOVA followed by Sidak's multiple comparisons test, different letters (a, b and c) between bars indicate P < 0.05

Article Snippet: Primary porcine skeletal muscle satellite cells were treated with 10 μmol/L LY294002, a PI3K inhibitor (MCE, HY-10108), or 1 μmol/L SCH772984, an ERK1/2 inhibitor (MCE, HY-50846).

Techniques: Protein-Protein interactions, RNA Sequencing, Knock-Out, Knockdown, Expressing, Analysis, Cell Culture, Two Tailed Test

Journal: Journal of Animal Science and Biotechnology

Article Title: ZBED6 deletion activates FNDC1 -mediated PI3K/AKT and MAPK/ERK1/2 signaling pathways to promote myogenesis in pigs

doi: 10.1186/s40104-026-01504-w

Figure Lengend Snippet: Inhibition of PI3K/AKT and MAPK/ERK1/2 pathways reduces the effects of the ZBED6-FNDC1 axis on myogenesis. A and B representative immunofluorescence staining for EdU ( A ) and percentage of EdU positive cells ( B ) in ZBED6 knockdown primary porcine skeletal muscle satellite cells treated with SCH772984, n = 3 independent cell-culture experiments. C and D . representative immunofluorescence staining for EdU ( C ) and percentage of EdU positive cells ( D ) in ZBED6 knockdown primary porcine skeletal muscle satellite cells treated with LY294002, n = 3 independent cell-culture experiments. E and F representative immunofluorescence staining for MYHC ( E ) and fusion index ( F ) in ZBED6 knockdown primary porcine skeletal muscle satellite cells treated with SCH772984, n = 3 independent cell-culture experiments. G and H representative immunofluorescence staining for MYHC ( G ) and fusion index ( H ) in ZBED6 knockdown primary porcine skeletal muscle satellite cells treated with LY294002, n = 3 independent cell-culture experiments. Data are represented as mean ± SEM. In (B), (D), (F), and (H), the P values were determined by two-way ANOVA followed by Sidak's multiple comparisons test, different letters (a and b) between bars indicate P < 0.05

Article Snippet: Primary porcine skeletal muscle satellite cells were treated with 10 μmol/L LY294002, a PI3K inhibitor (MCE, HY-10108), or 1 μmol/L SCH772984, an ERK1/2 inhibitor (MCE, HY-50846).

Techniques: Inhibition, Immunofluorescence, Staining, Knockdown, Cell Culture

Journal: MedComm

Article Title: TRIM28 Increases RFC4 Protein Expression to Promote Triple‐Negative Breast Cancer Progression

doi: 10.1002/mco2.71043

Figure Lengend Snippet: TRIM28 ubiquitinates RFC4 with K29‐linked polyubiquitin chains. (A) MDA‐MB‐231 cells were transduced with lentiviruses carrying empty vector or oeTRIM28 and treated with cycloheximide (CHX) for 0, 4, 8, and 12 h. (B) TRIM28 knockdown promoted RFC4 protein degradation through the proteasome pathway. (C) Analysis of the ubiquitin chain types on RFC4 affected by TRIM28. Ubiquitinated proteins were pulled down using anti‐Flag agarose under denaturing conditions from HEK293T cells transfected with the indicated constructs and analyzed by western blotting with the indicated antibodies. At 24 h after transfection, cells were treated with MG132 (10 µM) for an additional 4 h. (D) Analysis of endogenous RFC4 ubiquitination in MDA‐MB‐231 cells following siRNA‐mediated TRIM28 knockdown. (E) TRIM28‐mediated K29‐linked polyubiquitination of RFC4. (F) Effect of mutation of the TRIM28 enzymatic active site on the half‐life of RFC4. (G) Quantitative analysis of RFC4 protein levels after mutation of the TRIM28 enzymatic active site. (H) TRIM28 ubiquitinated RFC4 at K205 and K217. (I–K) Analysis of the half‐lives of Flag‐RFC4 wild‐type, K205R, K217R, and K205/217R mutant proteins in HEK293T cells.

Article Snippet: The proteasome inhibitor MG132, protein synthesis inhibitor CHX, autophagy inhibitor CQ and PI3K inhibitor LY294004 were purchased from MedChem Express (Shanghai, China).

Techniques: Transduction, Plasmid Preparation, Knockdown, Analysis, Ubiquitin Proteomics, Transfection, Construct, Western Blot, Mutagenesis

Journal: Advanced Science

Article Title: Early‐Life Manganese Overexposure Activates Microglial Phagocytosis through the YY1/TREM2 Axis Leading to Hippocampal Synaptic Remodeling in Mice

doi: 10.1002/advs.78036

Figure Lengend Snippet: Minocycline attenuates Mn‐induced synaptic remodeling and microglial phagocytic activation. (A) Schematic design map of animal experiment. (B) Representative 3D reconstruction showing microglial (IBA1, green) morphology. Scale bar, 30 µm. (C–E) Quantitative analysis of microglial morphology showing cell‐to‐body index, terminal points, and branch number under different treatment. (F) Quantitative analysis of microglial activation marker CD68 fluorescence intensity within IBA1 + microglia. (G,H) Representative immunofluorescence and 3D reconstruction of IBA1 + microglia phagocytosis of ES and IS pre‐ or postsynaptic markers (VGAT/PSD95), follow with quantification of phagocytosis levels within the microscopic field. Scale bar, 10 µm. (I,J) Representative Golgi–Cox stained CA1 neuronal dendritic, with quantification of filopodia, mushroom and total spine density ( n µm −1 ). Scale bar, 5 µm. (K,L) Representative fluorescence Z‐stack projection images of excitatory synaptic markers VGLUT1 (green) and PSD95 (red), and inhibitory synaptic markers VGAT (red) and Gephyrin (green), showing colocalization following Imaris‐based 3D reconstruction, together with Pearson's correlation coefficient analysis and quantification of the ES/IS ratio relative to control. Scale bar, 50 µm. Data are presented as mean ± SD from 4 mice per group. Data were analyzed using two‐way ANOVA followed by Tukey's multiple comparisons test, * p < 0.05, ** p < 0.01 versus Control group. # p < 0.05, ## p < 0.01 versus 1.1 mg kg −1 Mn group. See also Figure .

Article Snippet: In some cases, the microglial activation inhibitor minocycline (Mino; MCE, USA) was used in order to investigate whether Mn‐induced microglial activation led to synaptic remodeling.

Techniques: Activation Assay, Morphology, Analysis, Marker, Fluorescence, Immunofluorescence, Staining, Control