Journal: Aging Cell

Article Title: HNRNPC Succinylation Influences the Neurodegeneration of Alzheimer's Disease Through YME1L1 ‐Mediated Mitochondrial Metabolism

doi: 10.1111/acel.70646

Figure Lengend Snippet: YME1L1 promotes neuronal mitochondrial damage after AD. (A) CCK8 assay for cell viability. (B) Flow cytometry for apoptosis detection. (C) TEM for mitochondrial assessment (The green arrow indicates normal mitochondria, and the red arrow indicates damaged mitochondria). (D) ATP level measurement. (E) JC‐1 staining for cellular mitochondrial membrane potential measurement. (F) MitoSOX fluorescence staining for cellular mitochondrial ROS analysis. n = 3. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Intracellular superoxide levels were measured using the MitoSOX Red fluorescent probe (HY‐D1055; MCE, USA).

Techniques: CCK-8 Assay, Flow Cytometry, Staining, Membrane, Fluorescence

Journal: Aging Cell

Article Title: HNRNPC Succinylation Influences the Neurodegeneration of Alzheimer's Disease Through YME1L1 ‐Mediated Mitochondrial Metabolism

doi: 10.1111/acel.70646

Figure Lengend Snippet: YME1L1 induces neuronal mitochondrial damage and neuronal death following AD through OPA1 hydrolysis. (A) WB detection of OPA1 expression in hippocampal tissue ( n = 6). (B) WB detection of OPA1 expression. (C) CCK8 assay for cell viability. (D) Flow cytometry for apoptosis detection. (E) TEM for mitochondrial detection (The red arrow indicates damaged mitochondria). (F) ATP level detection. (G) JC‐1 staining for measuring mitochondrial membrane potential. (H) MitoSOX fluorescence staining for analyzing mitochondrial ROS. n = 3. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Intracellular superoxide levels were measured using the MitoSOX Red fluorescent probe (HY‐D1055; MCE, USA).

Techniques: Expressing, CCK-8 Assay, Flow Cytometry, Staining, Membrane, Fluorescence

Journal: Aging Cell

Article Title: HNRNPC Succinylation Influences the Neurodegeneration of Alzheimer's Disease Through YME1L1 ‐Mediated Mitochondrial Metabolism

doi: 10.1111/acel.70646

Figure Lengend Snippet: HNRNPC succinylation modifies neuronal mitochondrial damage and neuronal death following AD via YME1L1. (A) PCR and WB detection of YME1L1, WB detection of OPA1. (B) CCK8 analysis of cell activity. (C) Flow cytometry to detect cell apoptosis. (D) TEM detection of mitochondria (The green arrow points to normal mitochondria, and the red arrow points to damaged mitochondria). (E) ATP level detection. (F) JC‐1 staining was used to measure the mitochondrial membrane potential of cells. (G) MitoSOX fluorescent staining analysis of mitochondrial ROS in cells. n = 3. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Intracellular superoxide levels were measured using the MitoSOX Red fluorescent probe (HY‐D1055; MCE, USA).

Techniques: Activity Assay, Flow Cytometry, Staining, Membrane

Journal: Aging Cell

Article Title: HNRNPC Succinylation Influences the Neurodegeneration of Alzheimer's Disease Through YME1L1 ‐Mediated Mitochondrial Metabolism

doi: 10.1111/acel.70646

Figure Lengend Snippet: SIRT5 affects neuronal mitochondrial function by desuccinylating HNRNPC. (A) WB detection of YME1L1 and OPA1 expression. (B) CCK8 analysis of cell activity. (C) Flow cytometry detection of cell apoptosis. (D) TEM detection of mitochondrial morphology (The green arrow points to normal mitochondria, and the red arrow points to damaged mitochondria). (E) ATP level detection. (F) JC‐1 staining to measure cell mitochondrial membrane potential. (G) MitoSOX fluorescent staining analysis of mitochondrial ROS in cells. n = 3. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns stands for no significant difference.

Article Snippet: Intracellular superoxide levels were measured using the MitoSOX Red fluorescent probe (HY‐D1055; MCE, USA).

Techniques: Expressing, Activity Assay, Flow Cytometry, Staining, Membrane

Journal: Advanced Science

Article Title: Targeting Lactate‐Driven Stromal Autophagy via MCT1 Disrupts the Immunosuppressive Niche and Sensitizes Pancreatic Cancer to PD‐1 Blockade

doi: 10.1002/advs.76008

Figure Lengend Snippet: Lactate can induce activation of PSCs. (A) Representative microscopic images of primary mouse stellate cells under PBS, LAC 10 m m , LAC 15 m m , LAC 20 m m , and TGF‐β1 treatment conditions at 1 day, 3 days, and 5 days ( n = 5), scale bars 10 µm. (B) Fold change of per PSCs area in Figure . (C) Intracellular lactate concentration in PSCs ( n = 3). (D) Representative α‐SMA fluorescence staining images of primary mouse stellate cells treated with PBS, lactate or TGF‐β1 for 3 days ( n = 5), with nuclei stained by DAPI. Scale bars: 50 µm. α‐SMA positive area of individual PSCs. (E) Western blot analysis of α‐SMA, Collagen I, and FAP expression in the PSCs ( n = 3). (F) The mRNA levels of Collagen I, α‐SMA, FAP were measured by quantitative RTPCR assays ( n = 3). (G) Collagen I in the supernatant of PSCs culture medium was detected using the ELISA kit ( n = 3). (H) Intracellular TG concentration in PSCs. (I) Lipid droplets in PSC were stained with Oil Red O and observed under a light microscope, with nuclei stained by hematoxylin ( n = 3). Scale bars: 20 µm. Data are mean and ± SEM. *** p < 0.001. Statistical significance was determined by one‐way ANOVA or two‐sided Student's t‐test as appropriate.

Article Snippet: Cell count and viability was estimated using fluorescence Cell Analyzer (Countstar Rigel S2) with AO/PI reagent after removal erythrocytes (Miltenyi 130‐094‐183) and then debris and dead cells removal was decided to be performed or not (Miltenyi 130‐109‐398/130‐090‐101).

Techniques: Activation Assay, Concentration Assay, Fluorescence, Staining, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Light Microscopy

Journal: Advanced Science

Article Title: Targeting Lactate‐Driven Stromal Autophagy via MCT1 Disrupts the Immunosuppressive Niche and Sensitizes Pancreatic Cancer to PD‐1 Blockade

doi: 10.1002/advs.76008

Figure Lengend Snippet: MCT1 is a specific transporter mediating lactate‐induced PSCs activation. (A) Representative α‐SMA fluorescence staining images of primary mouse PSCs treated with si‐Scramble, si‐MCT4, si‐GPR132, si‐GPR81, or si‐MCT1 transfection under LAC 15 m m culture conditions for 3 days. Nuclei stained by DAPI. Scale bars: 50 µm. α‐SMA positive area of individual PSCs ( n = 5). (B) Western blot analysis of α‐SMA, Collagen I, and FAP expression in the PSCs ( n = 3). (C) The mRNA levels of α‐SMA and COL1A1 were measured by quantitative RT‐PCR assays ( n = 3). (D) Collagen I in the supernatant of the culture medium was detected using the ELISA kit ( n = 3). (E) Representative α‐SMA immunofluorescence staining images of primary mouse PSCs with or without lactate and AZD3965 treated for 3 days. Nuclei was stained by DAPI. Scale bars: 50 µm. α‐SMA positive area of individual PSCs ( n = 5). (F) The mRNA levels of α‐SMA and COL1A1 of PSCs were measured by quantitative RT‐PCR assays ( n = 3). (G) Western blot analysis of α‐SMA, Collagen I, and FAP expression in the PSCs ( n = 3). (H) Collagen I in the supernatant of PSCs culture medium detected by using an ELISA kit ( n = 3). (I) Single‐cell violin plot of MCT1 expression in human PC patients. Data are mean and ± SEM. *** p < 0.001; n.s., not significant. Statistical significance was determined by one‐way ANOVA or two‐sided Student's t‐test as appropriate.

Article Snippet: Cell count and viability was estimated using fluorescence Cell Analyzer (Countstar Rigel S2) with AO/PI reagent after removal erythrocytes (Miltenyi 130‐094‐183) and then debris and dead cells removal was decided to be performed or not (Miltenyi 130‐109‐398/130‐090‐101).

Techniques: Activation Assay, Fluorescence, Staining, Transfection, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Single Cell

Journal: Advanced Science

Article Title: Targeting Lactate‐Driven Stromal Autophagy via MCT1 Disrupts the Immunosuppressive Niche and Sensitizes Pancreatic Cancer to PD‐1 Blockade

doi: 10.1002/advs.76008

Figure Lengend Snippet: Lactate activates PSCs by inducing autophagy. (A) Representative confocal microscopy images of the red‐only puncta and the yellow puncta in PSCs ( n = 3). PSCs Nuclei was stained with DAPI. Scale bar: 20 µm. (B) Western blot analysis of LC3 I/II and p62 expression in PSCs ( n = 3). (C) Transmission electron microscopy of the autophagosome and autolysosomes of PSCs ( n = 3). Scale bars: 500 nm. Double arrow: autophagosome, single arrow: autolysosomes. (D) Representative fluorescence microscopy images of PSC after α‐SMA immunostaining. Scale bars: 10 µm. And α‐SMA positive area of individual PSC ( n = 5). (E) Collagen I in the supernatant of the PSCs culture medium detected by using an ELISA kit ( n = 3). Data are mean and ± SEM. *** p < 0.001. Statistical significance was determined by one‐way ANOVA or two‐sided Student's t‐test as appropriate.

Article Snippet: Cell count and viability was estimated using fluorescence Cell Analyzer (Countstar Rigel S2) with AO/PI reagent after removal erythrocytes (Miltenyi 130‐094‐183) and then debris and dead cells removal was decided to be performed or not (Miltenyi 130‐109‐398/130‐090‐101).

Techniques: Confocal Microscopy, Staining, Western Blot, Expressing, Transmission Assay, Electron Microscopy, Fluorescence, Microscopy, Immunostaining, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Targeting Lactate‐Driven Stromal Autophagy via MCT1 Disrupts the Immunosuppressive Niche and Sensitizes Pancreatic Cancer to PD‐1 Blockade

doi: 10.1002/advs.76008

Figure Lengend Snippet: Lactate induces autophagy in PSCs by regulating Vps34 lactylation. (A) Western blot analysis of pan‐Kla levels in PSC under normal condition, LAC 10 m m , LAC 15 m m , or LAC 20 m m for 3 days ( n = 3). (B) Western blot analysis of Vps34 and Pan‐Kla levels bound to Vps34 in PSCs ( n = 3). (C) Western blot analysis of ATG14, BECN1, Vps34, and the levels of pan‐Kla, ATG14, and BECN1 bound to Vps34 in PSCs with lactate treated or si‐MCT1 transfected ( n = 3). (D) Western blot analysis of ATG14, BECN1, Vps34, and the levels of pan‐Kla, ATG14 and BECN1 bound to Vps34 in PSCs with or without lactate and rapamycin treated, Vps34‐knockout‐sgRNA, Vps34‐knockout‐sgRNA + Vps34‐K 356/K781R , si‐ATG14, si‐BECN1 transfected ( n = 3). (E) Western blot analysis of Collagen I, α‐SMA, p62, and LC3 expression levels in PSCs with the above treated ( n = 3). (F) Representative fluorescence microscopy images of PSCs with the above treated ( n = 3) after α‐SMA immunostaining. Scale bars: 50 µm. (G) Representative images of lipid droplets in PSCs stained with Oil Red O, with nuclei stained by hematoxylin ( n = 3). Scale bar: 20 µm.

Article Snippet: Cell count and viability was estimated using fluorescence Cell Analyzer (Countstar Rigel S2) with AO/PI reagent after removal erythrocytes (Miltenyi 130‐094‐183) and then debris and dead cells removal was decided to be performed or not (Miltenyi 130‐109‐398/130‐090‐101).

Techniques: Western Blot, Transfection, Knock-Out, Expressing, Fluorescence, Microscopy, Immunostaining, Staining

Journal: Advanced Science

Article Title: Targeting Lactate‐Driven Stromal Autophagy via MCT1 Disrupts the Immunosuppressive Niche and Sensitizes Pancreatic Cancer to PD‐1 Blockade

doi: 10.1002/advs.76008

Figure Lengend Snippet: Development of orthotopic pancreatic cancer was impaired in PSC‐MCT1 −/− mice. (A) Gene Construction Strategy for COL1A2‐Cre‐ERT2‐MCT1 −/− Mice. (B) Double Immunofluorescence Staining of Collagen I and MCT1 in MCT1 −/− and MCT1 fl/fl mice. Nuclei are stained with DAPI. Scale bar: 20 µm. Relative MCT1 fluorescence intensity of Collagen I positive area. (C) Strategy for Constructing Conditional MCT1 −/− mice with PC in situ. Tamoxifen or corn oil is injected for 7 days, followed by the injection of KPC cells into the pancreas. In situ tumors were harvested 14 days later. (D–F) Representative Images of in situ tumors ( n = 4) in Figure (D), tumor weight statistics ( n = 5) (E), and double immunofluorescence observation of Collagen I and α‐SMA in tumors, with nuclei stained by DAPI ( n = 3) (F). Scale bar: 20 µm. (G) Diagram of the in vivo experimental regimen. (H–J) Tumor volumes (H) and representative Images of in situ tumors ( n = 5) (I) in Figure and tumor weight statistics ( n = 5) (J). Data are mean and ± SEM. *** p < 0.001; n.s., not significant. Statistical significance was determined by one‐way ANOVA or two‐sided Student's t‐test as appropriate.

Article Snippet: Cell count and viability was estimated using fluorescence Cell Analyzer (Countstar Rigel S2) with AO/PI reagent after removal erythrocytes (Miltenyi 130‐094‐183) and then debris and dead cells removal was decided to be performed or not (Miltenyi 130‐109‐398/130‐090‐101).

Techniques: Double Immunofluorescence Staining, Staining, Fluorescence, In Situ, Injection, Immunofluorescence, In Vivo

Journal: International Journal of Pharmaceutics: X

Article Title: Allicin-based biomimetic nanoparticles of the erythrocyte membrane for the delivery of lumefantrine to enhance its antimalarial effect

doi: 10.1016/j.ijpx.2026.100487

Figure Lengend Snippet: Neutralization of merozoites. A Purification of infected erythrocytes using the Percoll separation solution, displaying infected (black arrow) and normal (red arrow) erythrocytes. B Confocal fluorescent images of merozoites (blue), PECm-Allicin@LM (red), and their colocalization (purple). Scale bar = 50 μm. C Representative scatter plots of Hoechst 33342/CFDA-SE for the invasion test of merozoites and normal erythrocytes after drug treatment. D Intrusion rate (the percentage of cells in the Q2 area in each group relative to that in the Q2 area in the Model group). Data are presented as mean ± SEM ( n = 3), * P < 0.05. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The red fluorescent cell membrane dye DiD was purchased from Shanghai Titan Scientific Co., Ltd. (China).

Techniques: Neutralization, Purification, Infection