Journal: Translational Neurodegeneration

Article Title: Exercise promotes lipophagy and mitigates cognitive impairment in Alzheimer’s disease through downregulating microglial TMEM9

doi: 10.1186/s40035-026-00583-z

Figure Lengend Snippet: Tmem9 knockdown alleviates oligomeric Aβ (oAβ)-induced oxidative stress under conditions of glucose insufficiency in vitro. a , b Representative images and quantitative analysis of mitochondrial membrane potential (TMRE) and dihydroethidium (DHE) detecting reactive oxygen species in BV2 cells treated with or without oAβ under normal (25 mM glucose) or restricted (5 mM glucose) conditions. c, d Schematic of oleic acid (OA)-induced lipid droplet (LD) formation in BV2 cells, confirmed by Perilipin 2 immunostaining (Upper panel: 20 × objective, 5 × zoom; lower panel: 2 × magnification in the upper panel). e TMRE and DHE fluorescence staining in BV2 cells under glucose restriction (5 mM) following treatment with vehicle (Sham), oAβ alone, or OA+oAβ combination. f Quantification of TMRE and DHE fluorescence intensity as well as intracellular ATP levels in BV2 cells under glucose restriction (5 mM) with different treatments (Sham, oAβ, or OA+oAβ). g Experimental timeline for transfection of shRNA- Tmem9 or OE- Tmem9 lentiviral constructs, followed by OA+oAβ treatment in BV2 cells under glucose restriction conditions. h, i Effects of scramble control, shRNA- Tmem9 , negative control (NC), and OE- Tmem9 on TMRE and DHE fluorescence in OA+oAβ-treated BV2 cells under glucose restriction. j Schematic diagram depicting the proposed mechanism by which TMEM9 inhibits lipophagy-mediated ATP production to facilitate cellular senescence. Tmem9 knockdown and overexpression was implemented through plasmid transfection. Data are expressed as mean ± SD. n = 2 fields per well, 3 wells per group. One-way analysis of variance, followed by Tukey’s multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: The fluorescent dye tetramethylrhodamine ethyl ester (TMRE) (HY-D0985A, Storage concentration of 5 mM, MCE, NJ) was used to measure changes in mitochondrial membrane potential (ΔΨm).

Techniques: Knockdown, In Vitro, Analysis, Membrane, Immunostaining, Fluorescence, Staining, Transfection, shRNA, Construct, Control, Negative Control, Over Expression, Plasmid Preparation

Journal: Translational Neurodegeneration

Article Title: Exercise promotes lipophagy and mitigates cognitive impairment in Alzheimer’s disease through downregulating microglial TMEM9

doi: 10.1186/s40035-026-00583-z

Figure Lengend Snippet: Lipophagy protects microglia against oligomeric Aβ (oAβ)-induced oxidative stress under conditions of glucose insufficiency in BV2 microglial cells. a Experimental timeline for oleic acid (OA) and oAβ treatment combined with autophagy regulators rapamycin (Rap) and chloroquine (CQ) in BV2 cells under conditions of glucose insufficiency. b, c Representative fluorescence images and quantitative analysis of TMRE and DHE in sham, oAβ, OA+oAβ, OA+oAβ+Rap (autophagy inducer), OA+oAβ+CQ (autophagy inhibitor) groups, n =2 fields per well, 3 wells per group. d–g Western blots of autophagy markers (ATG5 and LC3II/LC3I ratio) among different treatment groups, n = 3 wells per group. h–m Western blots analysis of CPT1A, NDUFB10, and MTCO2 among different treatment groups, n = 3 wells per group. n–p Western blots analysis of ATPAF1, SDHB, IL-10, NLRP3, iNOS and IL-1β among these different treatment groups. Data are expressed as the mean ± SD, n = 3 wells per group. One-way analysis of variance, followed by Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The fluorescent dye tetramethylrhodamine ethyl ester (TMRE) (HY-D0985A, Storage concentration of 5 mM, MCE, NJ) was used to measure changes in mitochondrial membrane potential (ΔΨm).

Techniques: Fluorescence, Analysis, Western Blot

Journal: Translational Neurodegeneration

Article Title: Exercise promotes lipophagy and mitigates cognitive impairment in Alzheimer’s disease through downregulating microglial TMEM9

doi: 10.1186/s40035-026-00583-z

Figure Lengend Snippet: Tmem9 knockdown promotes lipophagy and maintains mitochondrial function through activation of lysosomal AMPKα signaling under condition of glucose insufficiency. a Western blotting analysis of phosphorylated AMPKα (p-AMPKα) and total AMPKα in cortical and hippocampal lysates from WT, Ctrl-5×FAD, PE-5×FAD, RNAi- Tmem9 , and PE+OE- Tmem9 mice, n = 4 male mice per group, aged 8 months. One-way analysis of variance (ANOVA), followed by Tukey’s multiple comparisons test. b Experimental timeline for oleic acid (OA) and oligomeric Aβ (oAβ) treatment in BV2 cells following Tmem9 knockdown (shRNA) or overexpression (OE). c Western blotting analysis of p-AMPKα/AMPKα in scramble control, shRNA- Tmem9 , negative control (NC), and OE- Tmem9 groups, n =3 wells per group. One-way analysis of variance (ANOVA), followed by Tukey’s multiple comparisons test. d Experimental design for OA+oAβ treatment in shRNA- Tmem9 -transfected BV2 cells treated with either DMSO or Compound C (an AMPK inhibitor) under glucose restriction conditions. e Effect of Compound C on AMPKα phosphorylation (p-AMPKα/AMPKα) in shRNA- Tmem9 BV2 cells. Statistical significance was assessed using independent-samples t -test. f Effect of Compound C on autophagy markers (ATG5, SQSTM1/P62, LC3II/LC3I) in DMSO- and Compound C-treated groups. g Effect of Compound C on the autophagy flux (autolysosome/autophagosome ratio) in DMSO- and Compound C-treated groups. h Representative images and quantification of TMRE and DHE following AMPK inhibition in the shRNA- Tmem9 cells following OA+oAβ treatment under condition of glucose insufficiency. Statistical significance was assessed using independent-samples t -test. Tmem9 knockdown was implemented through plasmid transfection. i-k Western blotting analysis of mitochondrial electron transport chain components (NDUFB10, UQCRC2, SDHB, MTCO2, ATPAF1), CPT1A, anti-inflammatory cytokine IL-10, and pro-inflammatory cytokines, including iNOS, NLRP3, IL-18, and IL-1β. Statistical significance was assessed using independent-samples t -test. Tmem9 knockdown and overexpression was implemented through lentivirus transfection. Data are expressed as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: The fluorescent dye tetramethylrhodamine ethyl ester (TMRE) (HY-D0985A, Storage concentration of 5 mM, MCE, NJ) was used to measure changes in mitochondrial membrane potential (ΔΨm).

Techniques: Knockdown, Activation Assay, Western Blot, Analysis, shRNA, Over Expression, Control, Negative Control, Transfection, Phospho-proteomics, Inhibition, Plasmid Preparation

Journal: Translational Neurodegeneration

Article Title: Exercise promotes lipophagy and mitigates cognitive impairment in Alzheimer’s disease through downregulating microglial TMEM9

doi: 10.1186/s40035-026-00583-z

Figure Lengend Snippet: Tmem9 downregulation promotes lysosomal AMPK activation through LKB1 translocation. a Co-immunoprecipitation demonstrating TMEM9–ATP6AP1 interaction in BV2 cells transfected with control (NC-Flag-Lv) or Tmem9 -overexpressing (OE- Tmem9 -Flag-Lv) vectors. b Experimental timeline for OA+oAβ treatment in BV2 cells transfected with control (NC) or OE- Tmem9 vectors, followed by DMSO or concanamycin A (Con A, V-ATPase inhibitor) treatment. c, d Representative images and quantitative analysis of TMRE and DHE mean intensities among NC+DMSO, NC+Con A, OE- Tmem9 +DMSO and OE- Tmem9 + Con A groups, n =2 fields per well, 3 wells per group. e–g Representative immunofluorescent images and quantitative analysis of LKB1 localization on LAMP1-positive lysosomal membranes, and LKB1-AXIN colocalization among Ctrl-5×FAD, PE-5×FAD, RNAi- Tmem9 , and PE+OE- Tmem9 groups, n = 4 male mice per group, at age of 8 months. h Proposed mechanistic diagram illustrating that exercise-induced activation of AMPK occurs through TMEM9-regulated lysosomal LKB1 translocation. Tmem9 overexpression in vitro was implemented through plasmid transfection. Data are expressed as mean±SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. One-way analysis of variance, followed by Tukey’s multiple comparisons test

Article Snippet: The fluorescent dye tetramethylrhodamine ethyl ester (TMRE) (HY-D0985A, Storage concentration of 5 mM, MCE, NJ) was used to measure changes in mitochondrial membrane potential (ΔΨm).

Techniques: Activation Assay, Translocation Assay, Co-Immunoprecipitation Assay, Transfection, Control, Analysis, Over Expression, In Vitro, Plasmid Preparation, Negative Control

Journal: iScience

Article Title: An experimental study on HRM combined with multiplex PCR for the identification of single and mixed diatom species

doi: 10.1016/j.isci.2026.117909

Figure Lengend Snippet:

Article Snippet: Eva Green Fluorescent Dye , Biotium , 31000.

Techniques:

Journal: bioRxiv

Article Title: Combined celecoxib and NDGA induces a non-canonical, autophagy-independent redox cell death via p62/KEAP1 in colorectal cancer

doi: 10.64898/2026.09.29.753356

Figure Lengend Snippet: A Representative flow cytometry plots showing global ROS levels measured by DCFDA staining in HCT116 and HT29 cells treated with the indicated compounds for 24 hours. The percentage of ROS positive cells is indicated within each plot. B Representative flow cytometry plots showing mitochondrial superoxide (O□•□) production measured by MitoSOX™ Green staining in HCT116 and HT29 cells treated with the indicated compounds for 24 hours. The percentage of MitoSOX positive cells is indicated within each plot. C Representative flow cytometry plots showing mitochondrial membrane potential (ΔΨm) measured by TMRM staining in HCT116 and HT29 cells treated with the indicated compounds for 24 hours. Red indicates cells with intact membrane potential, while green indicates cells with loss of ΔΨm. The percentage of cells in each quadrant is indicated within the plots. D Lipid peroxidation was measured using C11□BODIPY® 581/591 staining followed by flow cytometry in HCT116 and HT29 cells treated with the indicated compounds for 24 hours. Data are shown as mean ± SD (n=3). *** p = 0.0001; **** p < 0.0001 versus single agents (celecoxib or NDGA alone).

Article Snippet: Mitochondrial membrane potential was assessed using the fluorescent cationic dye Tetramethylrhodamine Methyl Ester (TMRM) (17109, TargetMol, Boston, MA, USA).

Techniques: Flow Cytometry, Staining, Membrane