Journal: Signal Transduction and Targeted Therapy

Article Title: BMX inhibition overcomes small cell lung cancer chemoresistance by stabilizing E2F1 via ERK1/2-Cyclin D1/CDK4/6 axis

doi: 10.1038/s41392-026-02644-1

Figure Lengend Snippet: Characterization of IHMT-15137 as a potent and selective BMX inhibitor. a Chemical structure of IHMT-15137 and its reversible analog IHMT-15138. b Determination of the IC 50 values of IHMT-15137, IHMT-15138 and ibrutinib against the BMX protein via the ADP-Glo kinase assay. c Anti-proliferative activity of IHMT-15137, IHMT-15138 and ibrutinib against TEL-BaF3-BMX cells determined by the CTG assay. d CETSA analysis of the effects of IHMT-15137 on BMX protein stability in H446DDPR and H69AR cell lysates: temperature- and dose-dependent stabilization of BMX by IHMT-15137. The right panels show the results of the quantitative analysis of the BMX protein levels from the Western blots. e DARTS assay confirmation of the direct binding of IHMT-15137 to BMX by assessing the stability of BMX against pronase-mediated proteolysis in H446DDPR and H69AR cell lysates. f Inhibitory effects of IHMT-15137 and IHMT-15138 on BMX phosphorylation (Tyr566) in TEL-BaF3-BMX cells after 4 h of treatment, as determined by Western blotting. g Inhibitory effects of 12 h of IHMT-15137 treatment on BMX phosphorylation (Tyr566) and its downstream signaling pathways in H446DDPR and H69AR cells. h Molecular docking analysis of the irreversible binding mode of IHMT-15137 with BMX (PDB ID: 3SXR). i Effect of IHMT-15137 on Biotin-PEG2-C2-iodoacetamide labeling of BMX via a competitive biotin-iodoacetamide assay. Data were presented as mean ± SEM ( n = 3)

Article Snippet: Subsequently, the samples were incubated with 8 mM Biotin‐PEG2‐C2‐iodoacetamide (TargetMol, Shanghai, China; Cat. No. T17565 ) for 4 h at room temperature in the dark.

Techniques: Kinase Assay, Activity Assay, CTG Assay, Western Blot, Binding Assay, Phospho-proteomics, Protein-Protein interactions, Labeling

Journal: The Journal of Biological Chemistry

Article Title: Starvation-induced HSC70 O-GlcNAcylation activates chaperone-mediated autophagy

doi: 10.1016/j.jbc.2026.113165

Figure Lengend Snippet: Glucose depletion enhances HSC70 O-GlcNAcylated at T430. A , the O-GlcNAcylation modification of endogenous HSC70 was detected under conditions of glucose deprivation or no treatment. B , the quantitation of ( A ). C , cells were transfected with HA-OGT and its O-GlcNAcylation was detected under conditions of glucose deprivation or no treatment. D , the quantitation of ( C ). E , cells were transfected with Flag-HSC70, then treated with 200 μmol/L Ac 3 6AzGlcNAc or not treated, and treated with 5 μmol/l TMG or not, as previously described . F , the quantitation of ( E ). G , HEK-293T cells were transfected with HA-HSC70-WT and HA-HSC70-T430A, and then the lysates were immunoprecipitated and immunoblotted with the antibodies indicated. H , the quantitation of ( G ). I , T430 of HSC70 is conserved in multiple species. The statistical analysis was performed using Student’s t test. ∗∗ p < 0.01; ∗∗∗∗ p < 0.0001. HSC70, heat shock cognate protein 70; O-GlcNAc, O-linked β-N-acetylglucosamine; OGT, O-GlcNAc transferase; TMG, Thiamet-G.

Article Snippet: The supernatant reacted with 50 μmol/L DBCO-PEG 4 -Biotin from TargetMol Chemicals Inc, 8 mmol/L urea, 10 mmol/L Hepes (pH 7.9) and Halt Protease & Phosphatase Inhibitor Cocktail (100 × ) from Thermo Fisher Scientific, then the pull-down complex isolated by streptavidin-coupled beads was subjected to Western blotting analysis.

Techniques: Modification, Quantitation Assay, Transfection, Immunoprecipitation, Analysis, Western Blot, Co-Immunoprecipitation Assay, Pull Down Assay

Journal: Molecular metabolism

Article Title: Epigenetic regulation of GABA catabolism in iPSC-derived neurons: The molecular links between FGF21 and histone methylation.

doi: 10.1016/j.molmet.2023.101798

Figure Lengend Snippet: Figure 2: Bulk RNA-seq using iPSC-derived forebrain neurons treated with FGF21, PF-05231023 and vehicle. (A) Volcano plots indicate differentially expressed genes with FDR 0.05. (B) Venn diagram showing expression profiles for the 974 genes with expression that could be affected by both FGF21 and PF-05231023 as determined by RNA-seq (FDR < 0.05). The middle and bottom Venn diagrams illustrate overlapping up-regulated and down-regulated genes in response to FGF21 and the long-acting FGF21 molecule. Those results indicate that those overlapping genes displayed identical directionality. (C) Heatmap showing expression profiles for the 974 genes affected by FGF21 and PF-05231023.

Article Snippet: Concentrations of the naïve form of FGF21 (Peprotech, cat#100-42) [22,23] and PF-05231023 (MedChemExpress, cat: HY-113697) [24,25] were selected to fall within the range of blood concentrations for these drugs observed during preclinical and clinical studies.

Techniques: RNA Sequencing, Derivative Assay, Expressing

Journal: Molecular metabolism

Article Title: Epigenetic regulation of GABA catabolism in iPSC-derived neurons: The molecular links between FGF21 and histone methylation.

doi: 10.1016/j.molmet.2023.101798

Figure Lengend Snippet: Figure 3: Plasma FGF21 concentrations were positively correlated with plasma GABA concentrations. (A) Plasma FGF21 concentrations were associated with plasma GABA levels in patients with alcohol use disorder. (B) both FGF21, and PF-05231023 could substantially increase the level of GABA in the culture medium of iPSC-derived forebrain neurons.

Article Snippet: Concentrations of the naïve form of FGF21 (Peprotech, cat#100-42) [22,23] and PF-05231023 (MedChemExpress, cat: HY-113697) [24,25] were selected to fall within the range of blood concentrations for these drugs observed during preclinical and clinical studies.

Techniques: Clinical Proteomics, Derivative Assay

Journal: Molecular metabolism

Article Title: Epigenetic regulation of GABA catabolism in iPSC-derived neurons: The molecular links between FGF21 and histone methylation.

doi: 10.1016/j.molmet.2023.101798

Figure Lengend Snippet: Figure 4: FGF21 and PF-05231023 induced GABA concentrations in iPSC-derived forebrain organoids. (A) A schematic outline of procedures used during the differentiation of iPSC-derived forebrain organoids. The middle panel displays images of iPSC-derived forebrain organoids during differentiation. The bottom panel of the schematic displays representative examples of staining for neural progenitors (SOX2) and neurons (TUJ1). (B) FGF21 and PF-05231023 substantially induced the concentrations of GABA in the culture medium of iPSC-derived forebrain organoids.

Article Snippet: Concentrations of the naïve form of FGF21 (Peprotech, cat#100-42) [22,23] and PF-05231023 (MedChemExpress, cat: HY-113697) [24,25] were selected to fall within the range of blood concentrations for these drugs observed during preclinical and clinical studies.

Techniques: Derivative Assay, Staining

Journal: Molecular metabolism

Article Title: Epigenetic regulation of GABA catabolism in iPSC-derived neurons: The molecular links between FGF21 and histone methylation.

doi: 10.1016/j.molmet.2023.101798

Figure Lengend Snippet: Figure 5: REST, EZH2 and H3K27me3 co-occupied the promoter regions of genes associated with GABA catabolism. (A) Heatmap showing expression profiles for the 15 genes involved in the GABAergic synapse pathway. (B) ChIP assays showing that REST, EZH2 and H3K27me3 co-occupied the promoter regions of the 13 selected genes as shown in Figure 5A that were associated with the GABAergic synapse pathway in response to FGF21 and PF-05231023 treatment of iPSC-derived forebrain neurons (n ¼ 4). Percentages of ChIP DNA/input were determined by qPCR. Data are represented as % input, (enrichment relative to IgG control) ¼ % input (antibody against REST, EZH2 and H3K27me3) e % input (IgG). ChIP-qPCR was performed in triplicate. Three independent experiments were performed. This figure represents results from one of the three experiments. (C) Co- Immunoprecipitation (IP) was used to determine whether REST protein could interact with EZH2 in iPSC-derived forebrain neurons. Whole cell lysates from iPSC-derived fore- brain neurons were immunoprecipitated with anti-REST antibodies or anti-IgG antibodies and protein samples were immunoblotted and probed with antibodies against REST or EZH2.

Article Snippet: Concentrations of the naïve form of FGF21 (Peprotech, cat#100-42) [22,23] and PF-05231023 (MedChemExpress, cat: HY-113697) [24,25] were selected to fall within the range of blood concentrations for these drugs observed during preclinical and clinical studies.

Techniques: Expressing, Derivative Assay, Control, ChIP-qPCR, Immunoprecipitation

Journal: Molecular metabolism

Article Title: Epigenetic regulation of GABA catabolism in iPSC-derived neurons: The molecular links between FGF21 and histone methylation.

doi: 10.1016/j.molmet.2023.101798

Figure Lengend Snippet: Figure 6: REST-EZH2-H3K37me3 complex-mediated gene repression in a drug dependent fashion. ChIP assays were performed in iPSC-derived forebrain neurons treated with FGF21, PF-05231023 and vehicle. We confirmed that changes in REST binding were correlated with changes in mRNA expression for all 13 genes. Specifically, in the presence of FGF21, all 13 binding sites except GNAI1 displayed significantly increased REST binding. In parallel, the recruitment of EZH2 and H3K27me3 also increased substantially, thus confirming that changes in REST-EZH2-H3K37me3 occupancy were highly correlated with transcription.

Article Snippet: Concentrations of the naïve form of FGF21 (Peprotech, cat#100-42) [22,23] and PF-05231023 (MedChemExpress, cat: HY-113697) [24,25] were selected to fall within the range of blood concentrations for these drugs observed during preclinical and clinical studies.

Techniques: Derivative Assay, Binding Assay, Expressing

Journal: Molecular metabolism

Article Title: Epigenetic regulation of GABA catabolism in iPSC-derived neurons: The molecular links between FGF21 and histone methylation.

doi: 10.1016/j.molmet.2023.101798

Figure Lengend Snippet: Figure 7: Schematic diagram illustrating the effects of FGF21 on GABA catabolism. Specifically, FGF21 and PF-05231023 significantly induced GABA levels in iPSC-derived forebrain neurons. We identified a drug-dependent occupancy of REST, EZH2, H3K27me3 in the promoter regions of genes associated with GABA catabolism i.e., ABAT (a key enzyme responsible for GABA catabolism), GRBBR1 (GABA B receptor 1), and SLC32A1 (GABA transporter), which resulted in transcriptional repression.

Article Snippet: Concentrations of the naïve form of FGF21 (Peprotech, cat#100-42) [22,23] and PF-05231023 (MedChemExpress, cat: HY-113697) [24,25] were selected to fall within the range of blood concentrations for these drugs observed during preclinical and clinical studies.

Techniques: Derivative Assay