Journal: Advanced Science

Article Title: Carrier‐Free Berberine/Nitidine Chloride Self‐Assembled Nanoparticles Induce Ferroptosis to Overcome Bortezomib Resistance in Multiple Myeloma

doi: 10.1002/advs.77174

Figure Lengend Snippet: Berberine (BBR)/nitidine chloride (NC) self‐assembled nanoparticles (BBR/NC‐SAPs) display potent and selective anti‐myeloma activity by inhibiting proliferation, clonogenicity, DNA synthesis, and inducing mitochondrial dysfunction. (A) Multiple myeloma cells (sensitive: 8226, KMS‐11; bortezomib‐resistant: 8226‐BTZR, KMS‐11‐BTZR) and normal B cells (GM12878) were treated for 48 h with a concentration gradient (0, 2, 4, 6, 8, and 10 µg/mL) of free BBR and NC combination (1:1). (B) Multiple myeloma cells (sensitive: 8226, KMS‐11; bortezomib‐resistant: 8226‐BTZR, KMS‐11‐BTZR) and normal B cells (GM12878) were treated for 48 h with a concentration gradient (0, 2, 4, 6, 8, and 10 µg/mL) of BBR/NC‐SAPs. (C) Cells were treated with the indicated concentrations (0, 2, 4, 6, 8, and 10 µg/mL) of BBR/NC‐SAPs. Colony formation assay demonstrating the potent, dose‐dependent inhibition of clonogenic survival. (D) Cells were treated with the indicated concentrations (0, 2, 4, 6, 8, and 10 µg/mL) of BBR/NC‐SAPs. Analysis of DNA synthesis by EdU incorporation assay utilizing flow cytometry. The percentage of EdU‐positive (green) cells decreased in a dose‐dependent manner upon treatment. (E) Assessment of mitochondrial membrane potential by JC‐1 staining employing flow cytometry. Treatment with BBR/NC‐SAPs induced a dose‐dependent loss of mitochondrial membrane potential. Data are presented as the mean ± SD from three independent experiments. Statistical significance was determined by a one‐way ANOVA followed by Dunnett's multiple comparison test versus the control group. * p < 0.05, ** p < 0.01.

Article Snippet: The mitochondrial membrane potential (ΔΨm) was assessed utilizing the fluorescent probe JC‐1 (MCE, USA) according to the manufacturer's protocol.

Techniques: Activity Assay, DNA Synthesis, Concentration Assay, Colony Assay, Inhibition, Flow Cytometry, Membrane, Staining, Comparison, Control

Journal: Advanced Science

Article Title: Carrier‐Free Berberine/Nitidine Chloride Self‐Assembled Nanoparticles Induce Ferroptosis to Overcome Bortezomib Resistance in Multiple Myeloma

doi: 10.1002/advs.77174

Figure Lengend Snippet: NR2F2 is essential for berberine (BBR)/nitidine chloride (NC) self‐assembled nanoparticles (BBR/NC‐SAPs)‐induced ferroptosis in multiple myeloma (MM) cells. (A) Cytotoxicity assays showing that NR2F2‐knockout cells locked the survival threshold and abolished the dosage‐dependent cytotoxicity of BBR/NC‐SAPs as compared to the control. (B) Colony formation assays revealing that NR2F2 knockout preserves clonogenic potential after BBR/NC‐SAPs treatment. (C) Analysis of DNA synthesis by EdU incorporation assay utilizing flow cytometry, demonstrating that NR2F2 knockout preserves cell proliferation potential after BBR/NC‐SAPs treatment. (D) Assessment of mitochondrial membrane potential (ΔΨm) via JC‐1 or similar dye, denoting that NR2F2 deletion prevents membrane depolarization triggered by BBR/NC‐SAPs. (E) Quantification of intracellular Fe 2+ showing blocked iron accumulation in NR2F2‐deficient cells upon BBR/NC‐SAPs treatment. (F) Lipid ROS levels measured by flow cytometry demonstrating that BBR/NC‐SAPs fail to elevate lipid peroxidation in NR2F2‐knockout cells. Ferroptosis‐related protein levels measured by western blot showing that BBR/NC‐SAPs fail to elevate lipid peroxidation in NR2F2‐knockout cells (G, 8226 and 8226‐BTZR; H, KMS‐11 and KMS‐11‐BTZR).

Article Snippet: The mitochondrial membrane potential (ΔΨm) was assessed utilizing the fluorescent probe JC‐1 (MCE, USA) according to the manufacturer's protocol.

Techniques: Knock-Out, Control, DNA Synthesis, Flow Cytometry, Membrane, Western Blot

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