Journal: Metabolic Brain Disease

Article Title: Adropin ameliorates behavioral seizures and the relevant neuroinflammation, oxidative stress, and neural damage in a rat model of pentylenetetrazole-induced seizure potentially by reducing the activation of NF-κB/IkB-α signaling pathway

doi: 10.1007/s11011-025-01654-2

Figure Lengend Snippet: Antioxidant and neuroprotective effects of adropin in the cortex in PTZ-induced epileptic conditions. Effects of adropin and other drug administrations on the levels of MDA ( A ), SOD1 ( B ), GFAP ( C ) and BDNF ( D ) in the cortex. AD1, 2 µg/kg dose of adropin; AD2, 10 µg/kg dose of adropin; LN, L-NAME; PTZ, pentylenetetrazole; VPA, valproic acid. * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: ELISA kits for rat interleukin (IL)−1β, TNF-α, brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP), malondialdehyde (MDA), and superoxide dismutase-1 (SOD1) were purchased from Elabscience (Houston, Texas, USA).

Techniques:

Journal: Metabolic Brain Disease

Article Title: Adropin ameliorates behavioral seizures and the relevant neuroinflammation, oxidative stress, and neural damage in a rat model of pentylenetetrazole-induced seizure potentially by reducing the activation of NF-κB/IkB-α signaling pathway

doi: 10.1007/s11011-025-01654-2

Figure Lengend Snippet: Antioxidant and neuroprotective effects of adropin in the hippocampus in PTZ-induced epileptic conditions. Effects of adropin and other drug administrations on the levels of MDA ( A ), SOD1 ( B ), GFAP ( C ) and BDNF ( D ) in the hippocampus. AD1, 2 µg/kg dose of adropin; AD2, 10 µg/kg dose of adropin; LN, L-NAME; PTZ, pentylenetetrazole; VPA, valproic acid. * p < 0.05, ** p < 0.01 and *** p < 0.001

Article Snippet: ELISA kits for rat interleukin (IL)−1β, TNF-α, brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP), malondialdehyde (MDA), and superoxide dismutase-1 (SOD1) were purchased from Elabscience (Houston, Texas, USA).

Techniques:

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Autophagy is activated in NSC-34 cells expressing ALS-linked misfolded proteins. (A) Immunoblot analysis of macroautophagy and CMA markers in NSC-34 cells stably transfected with wild-type (WT) or mutant G85R or G93A SOD1. Quantification of (B) LC3-II, (C) SQSTM1, (D) LAMP2A and (E) HSPA8 protein levels from immunoblots normalized to WT SOD1-expressing cells. (F) Immunoblot analysis of macroautophagy and CMA markers in NSC-34 cells stably transfected with WT or mutant Q331K or M337V TARDBP. Quantification of (G) LC3-II, (H) SQSTM1, (I) LAMP2A and (J) HSPA8 protein levels from immunoblots normalized to WT TARDBP-expressing cells. Data represent mean ± SD, n = 5-6 independent experiments, * p <0.05 and ** p <0.01 compared to cells expressing the WT form of relevant protein using one-way ANOVA with Tukey's posthoc test.

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Expressing, Western Blot, Stable Transfection, Transfection, Mutagenesis

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Rilmenidine stimulates macroautophagy and clears mutant SOD1 protein from NSC-34 cells. (A) Immunoblot analysis of macroautophagy markers in NSC-34 cells transiently transfected with wild-type (WT) or mutant SOD1 A4V and treated with rilmenidine (Ril) for 18 h post-transfection. Quantification of (B) LC3-II and (C) SQSTM1 protein levels from immunoblots normalized to untreated cells. Data represent mean ± SD, n = 4-6 independent experiments, * p <0.05 compared to untreated cells using one-way ANOVA with Tukey's post hoc test. (D) Photomicrographs of NSC-34 cells transfected with mCherry-GFP-LC3 plasmid and treated with rilmenidine. In rilmenidine-treated cells, there is a significantly increased proportion of mCherry-positive mature autolysosomes without GFP (arrows), indicating fusion of autophagosomes (AP) and lysosomes. (E) Quantification of the percentage of mCherry-positive mature autolysosomes relative to total puncta per cell. Data represent mean ± SD, n = 3 independent experiments, * p <0.05 using an unpaired t-test. (F) Immunoblot analysis of LC3-II levels in NSC-34 cells transfected with mutant SOD1 A4V and treated with 400 nM bafilomycin A 1 (Baf A1) ± 10 µM rilmenidine for 24 h. (G) Quantification of LC3-II protein levels from immunoblots normalized to cells treated with bafilomycin A 1 only. Data represent mean ± SD, n = 3 independent experiments, * p <0.05 compared to bafilomycin A 1 -treated cells using an unpaired t-test. (H) Immunoblot analysis of human SOD1 (HsSOD1) levels in WT or mutant SOD1 A4V -transfected NSC-34 cells treated with rilmenidine for 24 h post-transfection. (I) Quantification of HsSOD1 protein levels from immunoblots normalized to untreated cells. Data represent mean ± SD, n = 5 independent experiments, * p <0.05 compared to untreated cells using one-way ANOVA with Tukey's posthoc test. (J) Effect of rilmenidine or rotenone (positive control) on cell viability determined by MTT reduction assay (expressed as % of untreated cells). Data represent mean ± SD, n = 3 independent experiments. ** p <0.01, *** p <0.001 compared to untreated cells using one-way ANOVA with Tukey's posthoc test.

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Mutagenesis, Western Blot, Transfection, Plasmid Preparation, Positive Control, MTT Reduction Assay

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Rilmenidine has no effect on the CMA pathway in NSC-34 cells. (A) Immunoblot analysis of CMA markers and substrate SNCA in NSC-34 cells transiently transfected with wild-type (WT) SOD1 and treated with rilmenidine (Ril) for 18 h post-transfection. Quantification of (B) LAMP2A, (C) HSPA8 and (D) SNCA protein levels from immunoblots normalized to untreated cells. Data represent mean ± SD, n = 3 independent experiments.

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Western Blot, Transfection

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Rilmenidine stimulates macroautophagy in H9 human embryonic stem cell-derived spinal motor neurons. Photomicrographs of motor neuron spheres stained for (A) HB9 and (B) ISL1, and dissociated mature motor neurons stained with (C) CHAT and (D) TUBB3 antibodies. (E) Immunoblot analysis of macroautophagy markers in induced motor neurons transiently transfected with wild-type SOD1 and treated with rilmenidine (Ril) for 24 h post-transfection. Quantification of (F) LC3-II and (G) SQSTM1 protein levels from immunoblots normalized to untreated cells. Data represent mean ± SD, n = 5 independent experiments, * p <0.05 compared to untreated cells using an unpaired t-test.

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Derivative Assay, Staining, Western Blot, Transfection

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Rilmenidine treatment stimulates MTOR-independent macroautophagy and mitophagy in motor neurons of SOD1 G93A mice. (A) Immunoblot analysis of macroautophagy and CMA markers in lumbar spinal cords from vehicle- or rilmenidine-treated SOD1 G93A mice at 90 days of age. Quantification of (C) LC3-II, (D) SQSTM1 and (E) VDAC1 protein levels from immunoblots normalized to vehicle group. (B) Immunoblot analysis of autophagy markers and phosphorylated (p-MTOR) and total MTOR levels in spinal cords from vehicle- or rilmenidine-treated SOD1 G93A mice at end stage. Quantification of (C) LC3-II, (D) SQSTM1 and (E) VDAC1 protein levels and (F) p-MTOR:MTOR ratio from immunoblots normalized to vehicle group. Data represent mean ± SD, n = 3-5 mice, * p <0.05 compared to vehicle-treated mice using an unpaired t-test.

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Western Blot

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Rilmenidine treatment increases autophagosome number in spinal motor neurons of SOD1 G93A mice. (A) LC3 immunohistochemical analysis in lumbar spinal cords of vehicle- and rilmenidine-treated SOD1 G93A mice. In vehicle-treated mice, there is diffuse distribution of autophagosomes in the cytoplasm of RBFOX3/NeuN-positive motor neurons. In rilmenidine-treated mice, there is accumulation of autophagosomes (arrows) in motor neurons. (B) LAMP2 immunohistochemical analysis in lumbar spinal cords of vehicle- and rilmenidine-treated SOD1 G93A mice. In vehicle- and rilmenidine-treated mice, lysosomes (arrows) are distributed in the cytoplasm of motor neurons. Quantification of (C) LC3-positive autophagosomes and (D) LAMP2-positive lysosomes in motor neurons. Data represent mean ± SD, n = 3 mice per group, * p <0.05 compared to vehicle-treated mice using an unpaired t-test.

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Immunohistochemical staining

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Rilmenidine treatment worsens disease progression and neurodegeneration in SOD1 G93A mice. (A) Onset of body weight decline and (B) locomotor activity were similar in rilmenidine- and vehicle-treated SOD1 G93A mice. (C) Survival was significantly reduced by rilmenidine treatment compared to vehicle-treated animals. Data represent mean ± SD, n = 10 mice per group, * p <0.05 using the log-rank test. (D) Photomicrographs of ventral horns stained with cresyl violet in lumbar spinal cords of mice at 90 days of age. (E) Motor neuron counts in ventral horns of lumbar spinal cords of vehicle- and rilmenidine-treated mice. Motor neuron numbers in rilmenidine-treated mice were significantly decreased compared to vehicle-treated mice. Data represent mean ± SD, n = 5 mice per group, ** p <0.01 compared to vehicle-treated mice using an unpaired t-test. Immunohistochemical analysis of (F) astrocytes using GFAP and (G) microglia using ITGAM/CD11b in lumbar spinal cords of mice at end stage. Astrocyte and microglial activation appear similar in spinal cords of rilmenidine- and vehicle-treated mice. Images are representative of 3 mice per group.

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Biomarker Discovery, Activity Assay, Staining, Immunohistochemical staining, Activation Assay

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Rilmenidine treatment increases the burden of pathological and aggregated SOD1 in motor neurons of SOD1 G93A mice. (A) Immunoblot analysis of soluble SOD1 protein levels in spinal cords of vehicle- and rilmenidine-treated SOD1 G93A mice at end stage. Quantification of (B) human SOD1 (HsSOD1) and (C) misfolded SOD1 (misSOD1) protein levels normalized to vehicle-treated mice. (D) Immunoblot analysis of insoluble SOD1 protein levels in spinal cord pellet fractions of vehicle- and rilmenidine-treated SOD1 G93A mice at end stage. (E) Quantification of HsSOD1 protein level in pellet fraction normalized to vehicle group. Data represent mean ± SD, n = 5 mice per group, ** p <0.01 compared to vehicle-treated mice using an unpaired t-test. (F, H and I) Misfolded SOD1 immunohistochemical analysis of lumbar spinal cords of vehicle- and rilmenidine-treated SOD1 G93A mice at 90 days of age. (F-H) In vehicle-treated mice, misfolded SOD1 shows diffuse staining in the cytoplasm of RBFOX3/NeuN-positive motor neurons (arrows). In rilmenidine-treated mice, SOD1 immunoreactivity is increased with accumulation of SOD1 into cytoplasmic aggregates in motor neurons. SOD1 accumulates into (H) Lewy body-like or (I) skein-like inclusions (arrows) in motor neurons. (G) Quantification of misfolded SOD1 aggregates in motor neurons. Data represent mean ± SD, n = 5 mice per group, ** p <0.01 compared to vehicle-treated mice using an unpaired t-test.

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Western Blot, Immunohistochemical staining, Staining

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Rilmenidine-induced SOD1 inclusions accumulate outside the autophagy system in motor neurons of SOD1 G93A mice. (A) Misfolded SOD1 and LC3 immunohistochemical analysis in lumbar spinal cords of vehicle- and rilmenidine-treated SOD1 G93A mice at 90 days of age. Vehicle-treated mice show colocalization of SOD1 and autophagosomes (arrowheads). Large SOD1 aggregates in rilmenidine-treated mice (arrows) do not colocalize with autophagosomes (arrowheads). (B) Misfolded SOD1 and LAMP2 immunohistochemical analysis in lumbar spinal cords of vehicle- and rilmenidine-treated SOD1 G93A mice at 90 days of age. Vehicle-treated mice show colocalization of SOD1 and lysosomes (arrowheads). Large SOD1 aggregates in rilmenidine-treated mice (arrows) do not colocalize with lysosomes (arrowheads).

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Immunohistochemical staining

Journal: Autophagy

Article Title: Rilmenidine promotes MTOR-independent autophagy in the mutant SOD1 mouse model of amyotrophic lateral sclerosis without slowing disease progression

doi: 10.1080/15548627.2017.1385674

Figure Lengend Snippet: Rilmendine induces excessive mitophagy and mitochondrial depletion in spinal motor neurons of SOD1 G93A mice. (A) Photomicrographs of NSC-34 cells transfected with mt-Rosella plasmid and treated with CCCP (positive control) or rilmenidine. In CCCP- and rilmenidine-treated cells, there is a significantly increased proportion of dsRed-positive puncta without pHluorin (arrowheads), indicating fusion of mitochondria and lysosomes. (B) Quantification of the percentage of dsRed-positive cells relative to total transfected cells. Data represent mean ± SD, n = 2 independent experiments, **p<0.01 and *** p <0.001 using a one-way ANOVA. (C) TOMM20 immunohistochemical analysis in lumbar spinal cords of vehicle- and rilmenidine-treated SOD1 G93A mice. (D) Quantification of TOMM20 immunoreactivity showing mitochondrial depletion in RBFOX3/NeuN-positive motor neurons of rilmenidine-treated mice Data represent mean ± SD, n = 4 mice per group, * p <0.05 compared to vehicle-treated mice using an unpaired t-test.

Article Snippet: Membranes were blocked with 5% (w:v) skim milk dried powder (Fonterra, 492281) in Tris-buffered saline with Tween-20 (TBST; Tris base 20 mM, NaCl 140 mM, pH 8.0, 0.1% [v:v] Tween-20 [Sigma, P7949]) for 30 min and incubated with antibodies to rabbit LC3B (1:1,000; Sigma-Aldrich, L7543), mouse SQSTM1 (1:500; Abcam, ab56416), sheep SOD1 (1:4,000; Merck, 574597), rabbit LAMP2A (1:500; Abcam, ab18528), mouse HSPA8/HSC70 (1:500; Enzo Lifesciences, ALX-804-067), mouse SNCA (1:1,000; BD Transduction Labs, 610786), rabbit VDAC1 (1:1,000; Abcam ab15895), rabbit MTOR (1:1000; Cell Signaling Technology, 2983), rabbit phospho-MTOR (1:1000; Cell Signaling Technology, 2971), mouse human-specific SOD1 (1:2,000; R&D Systems, MAB3418), mouse misfolded SOD1 (C4F6) (1:250; Medimabs, MM-0070-2-P) or mouse ACTB (1:2,000; Sigma-Aldrich, A5316) antibodies in 3% (w:v) BSA (Sigma-Aldrich, A3912) in TBST overnight at 4°C.

Techniques: Transfection, Plasmid Preparation, Positive Control, Immunohistochemical staining

Journal: bioRxiv

Article Title: Proteomic profiling reveals pleiotropic antimetabolite activity of triciribine in acute lymphoblastic leukemia

doi: 10.64898/2026.03.13.710465

Figure Lengend Snippet: A , UMAP visualization of drug sensitivity profiles for 528 compounds tested across 43 ALL cell lines. Each point represents a compound, positioned based on similarity of killing profiles. B , sDSS values for triciribine and PI3K-Akt-mTOR pathway inhibitors across 43 ALL cell lines. The dashed line indicates sDSS = 8. The mean sDSS value for each inhibitor is indicated by an ‘×’ or a larger dot. Akt inhibitors are indicated in red. C , Pairwise Spearman correlation analysis of drug sensitivity profiles for PI3K-Akt-mTOR pathway inhibitors across 43 ALL cell lines. Akt inhibitors are indicated in red. D , Immunoblot analysis of phosphorylated Akt (Ser473), total Akt, and the loading control SOD1 in ALL-SIL and P12-ICHIKAWA cells following treatment with indicated concentrations or durations of triciribine.

Article Snippet: Primary antibodies included ADK (AM8619b-ev; Nordic Biosite), SOD1 (sc-17767; Santa Cruz Biotechnology), PARP (9532), Akt (4691), and phospho-Akt (Ser473; 4060) (Cell Signaling Technology).

Techniques: Western Blot, Control

Journal: Animal Models and Experimental Medicine

Article Title: Adropin modulates pancreatic cell proliferation and glutathione levels in an animal model of type 1 diabetes mellitus

doi: 10.1002/ame2.70092

Figure Lengend Snippet: Impact of adropin on superoxide dismutase expression in pancreatic β‐cells of normoglycemic and diabetic rats. (A) Immunofluorescence labeling with anti‐superoxide dismutase antibody and anti‐insulin antibodies showed expression of superoxide dismutase in pancreatic β‐cells. (B) Quantification of the histological analysis showed a significant (*** p < 0.001) decrease in superoxide dismutase distribution in pancreatic endocrine cells and a significant (**** p < 0.0001) decrease in its localization in β‐cells of rats with diabetes when compared to the normal group. n = 6. Scale bar: 50 μm. Data analysis was done using the ANOVA test.

Article Snippet: Superoxide dismutase (rabbit) (1:500) , Rockland Immunochemicals, USA.

Techniques: Expressing, Immunofluorescence, Labeling

Journal: Animal Models and Experimental Medicine

Article Title: Adropin modulates pancreatic cell proliferation and glutathione levels in an animal model of type 1 diabetes mellitus

doi: 10.1002/ame2.70092

Figure Lengend Snippet: Effect of adropin on superoxide dismutase expression in pancreatic α‐cells of normal and diabetic rats. (A) Immunofluorescence staining using anti‐superoxide dismutase and anti‐glucagon antibodies showed expression of superoxide dismutase in pancreatic α‐cells. (B) Quantification of the histological analysis showed a significant (** p < 0.01) decrease in superoxide dismutase distribution in the pancreatic endocrine cells of diabetic rats compared to the normal group. α‐Cell expression of superoxide dismutase did not change with adropin treatment among all groups. n = 6. Scale bar: 50 μm. Data analysis was done using the ANOVA test.

Article Snippet: Superoxide dismutase (rabbit) (1:500) , Rockland Immunochemicals, USA.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Animal Models and Experimental Medicine

Article Title: Adropin modulates pancreatic cell proliferation and glutathione levels in an animal model of type 1 diabetes mellitus

doi: 10.1002/ame2.70092

Figure Lengend Snippet: Effect of adropin on catalase, superoxide dismutase and total glutathione activities in the serum samples of healthy and diabetic rats. (A) Catalase activity was significantly decreased in diabetic rats compared to normal controls. Adropin slightly increased catalase in DMT group. (B) Superoxide dismutase was slightly increased in DMT compared to the diabetic treated with adropin. (C) Total glutathione was significantly raised with adropin treatment in DMT compared to DMUT. n = 4–6. Data analysis was done using the ANOVA test. * p < 0.05.

Article Snippet: Superoxide dismutase (rabbit) (1:500) , Rockland Immunochemicals, USA.

Techniques: Activity Assay

Journal: Current Issues in Molecular Biology

Article Title: The Effects of Engeletin on Insulin Resistance Induced in Human HepG2 Liver Cells

doi: 10.3390/cimb47070535

Figure Lengend Snippet: ( A ) . The analysis results of the effect of Engeletin on MDA, ( B ) GSH, and ( C ) SOD (ENG: Engeletin; IR: insulin resistance; MET: metformin). *: p < 0.05.

Article Snippet: Oxidant and antioxidant parameters—malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) levels—were measured using human-specific ELISA kits (Wuhan, China) (MDA: ELABSCIENCE (E-BC-K025-M); GSH: ELABSCIENCE(E-EL-H5410); and SOD: ELABSCIENCE (E-EL-H1113)) with two repetitions.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Multitarget mechanisms of the herb pair Achyranthes bidentata and Paeonia lactiflora Pall. in ameliorating hypertensive cardiomyopathy: combining network pharmacology and functional exploration

doi: 10.3389/fphar.2026.1717533

Figure Lengend Snippet: Effects of AB-PL on oxidative stress and the Nrf2/HO-1 signaling pathway in the myocardial tissues of hypertensive mice: (A) reactive oxygen species (ROS) level; (B) Nrf2 and (C) HO-1 mRNA expression; (D) superoxide dismutase (SOD) enzyme activity; (E) Keap1, and (F) SOD protein expression. Immunohistochemical staining of (G) Nrf2 and (H) HO-1 (n ≥ 3; # p < 0.05, ## p < 0.01 vs. model group; * p < 0.05, ** p < 0.01 vs. control group).

Article Snippet: The proteins were subjected to SDS-PAGE electrophoresis, transferred to NC membranes, blocked with 5% skimmed milk for 2 h, and incubated overnight at 4 °C with nuclear factor erythroid-2-related factor 2 (Nrf2; Zen bioscience, China), Keap1 (Proteintech, United States), SOD (Proteintech, United States), heme oxygenase-1 (HO-1; Proteintech, United States), NLRP3 (Proteintech, United States), ASC (Proteintech, United States), and caspase-1 (Proteintech, United States).

Techniques: Expressing, Activity Assay, Immunohistochemical staining, Staining, Control