Journal: CNS Neuroscience & Therapeutics

Article Title: Microbial lipopolysaccharide‐induced inflammation contributes to cognitive impairment and white matter lesion progression in diet‐induced obese mice with chronic cerebral hypoperfusion

doi: 10.1111/cns.14301

Figure Lengend Snippet: High‐fat diet feeding increases lipopolysaccharide (LPS) concentration, inflammatory cytokine concentrations, TLR4 expression, and blood–brain barrier (BBB) permeability in the corpus callosum of wild‐type mice 14 and 28 days after bilateral carotid artery stenosis. (A) Western blotting and densitometric analyses of LPS and TLR4 ( n = 6 per group). (B) IL‐6 ( n = 5 per group) and IL‐1β levels ( n = 6 per group). (C) Western blotting and densitometric analysis of occludin ( n = 6 per group). (D) BBB permeability as assessed by IgG ( n = 6 per group). Scale bar: 50 μm. Results are presented as mean ± SD. * p < 0.05, ** p < 0.01.

Article Snippet: The membranes were blocked in Block Ace (Dainichi‐Seiyaku) and then incubated overnight at 4°C with antibodies against myelin basic protein (MBP; 1:5000; Abcam), LPS (1:5000; Abcam), TLR4 (1:5000; Santa Cruz Biotechnology), occludin (1:5000; Abcam), or α‐tubulin (1:10000; Santa Cruz Biotechnology), followed by incubation with peroxidase‐conjugated secondary antibodies (1:5000; Santa Cruz Biotechnology).

Techniques: Concentration Assay, Expressing, Permeability, Western Blot

Journal: bioRxiv

Article Title: Sigma-1 Receptor Promotes Glycolysis in Neuronal Systems by Suppressing GRIM19

doi: 10.1101/2025.07.28.667250

Figure Lengend Snippet: A-Western blot analysis of S1R expression and total protein staining in N2a cell protein extract. B (left)-N2a cell extra-cellular acidification rate (ECAR) measurement with the application of Rotenone and Antimycin A (Rot/AA) and 2-Deoxyglucose (2-DG); (right)-Simultaneous measurement of N2a cell oxygen consumption rate (OCR). C (from left to right)- Basal glycolysis; % proton efflux rate (PER) from glycolysis; Basal oxygen consumption; MitoOCR/glycoPER. D (left)-N2a cell extra-cellular acidification rate (ECAR) measurement with the application of Oligomycin (Oligom.), Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) and Rotenone and Antimycin A (Rot/AA); (right)-Simultaneous measurement of N2a cells oxygen consumption rate (OCR); E (from left to right)-Non mitochondrial oxygen consumption; Basal respiration; Maximal respiration; Proton leak; ATP production; Spare respiratory capacity; Coupling efficiency; Basal acidification rate. Statistical analysis: Student’s t-test (*p < 0.05,***p < 0.001). B-E Wt N2a in white, S1R KO N2a in red.

Article Snippet: Expanded cells were collected and cell lysates were analyzed for S1R protein expression by Western blot using the Santa Cruz Biotechnology B5 anti-S1R receptor antibody (sc-137075).

Techniques: Western Blot, Expressing, Staining

Journal: bioRxiv

Article Title: Sigma-1 Receptor Promotes Glycolysis in Neuronal Systems by Suppressing GRIM19

doi: 10.1101/2025.07.28.667250

Figure Lengend Snippet: A-hexokinase. B-PKM. C-GAPDH. D-Enolase. E-GRIM19. F-SDHA. G-UQCRC2. H-MTCO1. I-ATP5a. J-S1R. A to I (top) Relative signal intensity compared to Wt (bottom) Western blot and total protein staining. Statistical analysis A to I, Student’s t-test (*p < 0.05, ***p < 0.001).

Article Snippet: Expanded cells were collected and cell lysates were analyzed for S1R protein expression by Western blot using the Santa Cruz Biotechnology B5 anti-S1R receptor antibody (sc-137075).

Techniques: Western Blot, Staining

Journal: bioRxiv

Article Title: Sigma-1 Receptor Promotes Glycolysis in Neuronal Systems by Suppressing GRIM19

doi: 10.1101/2025.07.28.667250

Figure Lengend Snippet: A- Transiently transfected N2a cells (up : KO GFP, down: KO S1RGFP). B-%PER glycolysis. C-MitoOCR/glycoPER. D- Percentage of positive cells. E-Stabilized S1R-overexpressing N2a cells (up : KO GFP, down : KO S1RGFP);. F-%PER glycolysis. G-MitoOCR/glycoPER. H- Percentage of positive cells. I- (top) Enolase signal intensity compared to Wt; (bottom) Western blot and total protein staining. J- (top) GRIM19 signal intensity compared to Wt; (bottom) Western blot and total protein staining. K- (top) PDH signal intensity compared to Wt; (bottom) Western blot and total protein staining. L- (top) LDH signal intensity compared to Wt; (bottom) Western blot and total protein staining. Statistical analysis B, C, F, G, I, J, K and L One-way ANOVA followed by multiple comparisons (*p < 0.05, **p < 0.01, ***p < 0.001)

Article Snippet: Expanded cells were collected and cell lysates were analyzed for S1R protein expression by Western blot using the Santa Cruz Biotechnology B5 anti-S1R receptor antibody (sc-137075).

Techniques: Transfection, Western Blot, Staining

Journal: bioRxiv

Article Title: Sigma-1 Receptor Promotes Glycolysis in Neuronal Systems by Suppressing GRIM19

doi: 10.1101/2025.07.28.667250

Figure Lengend Snippet: A (left)-Cortical neurons extra-cellular acidification rate (ECAR) measurement with the application of Rotenone and Antimycin A (Rot/AA) and 2-Deoxyglucose (2-DG); A (right)-Simultaneous measurement of cortical neurons oxygen consumption rate (OCR); Neurons from Wt mice in white. Neurons from S1R KO mice in red. B (from left to right)- Basal glycolysis; % proton efflux rate (PER) from glycolysis; Basal oxygen consumption; MitoOCR/glycoPER. C-(top) Enolase signal intensity compared to Wt; (bottom) Western blot and total protein staining. D (left)-Cortical neuronal cell extra-cellular acidification rate (ECAR) measurement with the application of Oligomycin (Oligom.), Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) and Rotenone and Antimycin A (Rot/AA); D (right)-Simultaneous measurement of cortical neurons oxygen consumption rate (OCR); Neurons from Wt mice in white. Neurons from S1R KO mice in red. E (from left to right)-Non mitochondrial oxygen consumption; Basal respiration; Maximal respiration; Proton leak; ATP production; Spare respiratory capacity; Coupling efficiency; Basal acidification rate. F-(top) GRIM19 signal intensity compared to Wt; (Bottom) Western blot and total protein staining. Statistical analysis: Student’s t-test (*p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: Expanded cells were collected and cell lysates were analyzed for S1R protein expression by Western blot using the Santa Cruz Biotechnology B5 anti-S1R receptor antibody (sc-137075).

Techniques: Western Blot, Staining

Journal: bioRxiv

Article Title: Sigma-1 Receptor Promotes Glycolysis in Neuronal Systems by Suppressing GRIM19

doi: 10.1101/2025.07.28.667250

Figure Lengend Snippet: A-Wt and S1R KO N2a cell ratio NAD + /NADH measured with NAD/NADH-Glo. B-Wt and S1R KO N2a cells transfected with either GFP or S1RGFP plasmid, ratio of NAD + /NADH measured with NAD/NADH-Glo; C-Wt and S1R KO primary culture of cortical neuron ratio of NAD + /NADH measured with NAD/NADH-Glo. D-Pictures representing Wt N2a cells expressing Peredox with different concentrations of extracellular Lactate and Pyruvate. Green channel represents the Peredox sensor binding NADH and red channel represents the total amount of Peredox expressed in the cell. The ratio of green/ged represents the amount of Peredox binding NADH over the total quantity of Peredox in the cell. E-Representation of the Green/Red ratio over time with the application of a different ratio of [Lac]/[Pyr]. Wt N2a in white. S1R KO N2a in red. F- Dose response of green/red ratio over the [Lac]/[Pyr]. G- Cytosolic NADH/NAD + of Wt and S1R KO N2a cells. Statistical analysis: Student’s t-test (**p < 0.01, ***p < 0.001).

Article Snippet: Expanded cells were collected and cell lysates were analyzed for S1R protein expression by Western blot using the Santa Cruz Biotechnology B5 anti-S1R receptor antibody (sc-137075).

Techniques: Transfection, Plasmid Preparation, Expressing, Binding Assay

Journal: bioRxiv

Article Title: Sigma-1 Receptor Promotes Glycolysis in Neuronal Systems by Suppressing GRIM19

doi: 10.1101/2025.07.28.667250

Figure Lengend Snippet: A-Wt and S1R KO N2a cell glucose uptake measured with Glucose Uptake-Glo. B-Wt and S1R KO N2a cells transfected with either GFP or S1RGFP plasmid; glucose uptake measured with Glucose Uptake-Glo, C-Wt and S1R KO primary culture of cortical neuron glucose uptake measured with Glucose Uptake-Glo, D-PET-Scan 18 FDG experimental design. E (right)- PET-Scan 18 FDG results, Standardized uptake value (SUV) per brain area; E (left) 2-way ANOVA for each time point. Statistical analysis for A-C: Student’s t-test (*p < 0.05, ***p < 0.001), for E: 2-way ANOVA (****p < 0.0001).

Article Snippet: Expanded cells were collected and cell lysates were analyzed for S1R protein expression by Western blot using the Santa Cruz Biotechnology B5 anti-S1R receptor antibody (sc-137075).

Techniques: Transfection, Plasmid Preparation

Journal: bioRxiv

Article Title: Sigma-1 Receptor Promotes Glycolysis in Neuronal Systems by Suppressing GRIM19

doi: 10.1101/2025.07.28.667250

Figure Lengend Snippet: A- Transfected N2a cells (up : S1R KO tGFP, down : S1R KO IP3tGFP); B- Percentage of GFP-positive cells C-Ip3R3 signal intensity compared to Wt tGFP; D-%PER glycolysis; E-MitoOCR/glycoPER; F- Basal respiration; G-Maximal respiration; H- (left) GRIM19 signal intensity compared to Wt tGFP; (right) Western blot and total protein staining; I-Transfected N2a cells (up : S1R KO scramble, down : S1R KO GRIM19 KD); J- Percentage of GFP-positive cells; K-GRIM19 signal intensity compared to Wt ; L-Basal glycolysis; M-MitoOCR/glycoPER. Statistical analysis, One-way ANOVA followed by multiple comparisons (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001)

Article Snippet: Expanded cells were collected and cell lysates were analyzed for S1R protein expression by Western blot using the Santa Cruz Biotechnology B5 anti-S1R receptor antibody (sc-137075).

Techniques: Transfection, Western Blot, Staining

Journal: bioRxiv

Article Title: Sigma-1 Receptor Promotes Glycolysis in Neuronal Systems by Suppressing GRIM19

doi: 10.1101/2025.07.28.667250

Figure Lengend Snippet: Schema highlighting the connection between mitochondrial processes and glycolysis through the NADH/NAD ratio. The role of Complex I of the oxidative phosphorylation chain is included. Summary of the consequences of S1R deletion on the overall energy metabolism and Complex I.

Article Snippet: Expanded cells were collected and cell lysates were analyzed for S1R protein expression by Western blot using the Santa Cruz Biotechnology B5 anti-S1R receptor antibody (sc-137075).

Techniques: Phospho-proteomics