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

Journal: Cell Reports

Article Title: PD-1 high CXCR5 – CD4 + peripheral helper T cells promote CXCR3 + plasmablasts in human acute viral infection

doi: 10.1016/j.celrep.2022.111895

Figure Lengend Snippet:

Article Snippet: LPS from E. Coli, TLR4 ligand , NOVUS Biologicals , NBP2-25295.

Techniques: Purification, Recombinant, Cell Isolation, Enzyme-linked Immunosorbent Assay, DNA Library Preparation, Software

Journal: International Journal of Molecular Sciences

Article Title: Altered Intestinal Morphology and Microbiota Composition in the Autism Spectrum Disorders Associated SHANK3 Mouse Model

doi: 10.3390/ijms20092134

Figure Lengend Snippet: Altered gut morphology in Shank3αβ knock-out (KO) mice. ( A – D ) Histological evaluation of GI tract from wild type and Shank3αβ KO mice. ( A ) Longitudinal cross sections of Shank3αβ KO mice and wild type (WT) mice were stained with hematoxylin/eosin (HE) (upper panels) and periodic acid schiff (PAS) reaction (lower panels). Exemplary images are shown. ( B – D ) Morphological analysis of ( B ) villi length and ( C ) width, and ( D ) crypt depth reveals a significantly decreased villi length (Mann-Whitney U -test, p = 0.009; n = 5 animals per group) but not width ( p = 0.534), and normal crypt depth ( p = 0.983) in Shank3αβ KO mice. ( E , F ) Immunohistochemistry was performed on 5 mice per group and 5 optic fields of view each from 3 sections per mouse were analyzed. ( E ) A slight but non-significant decrease in FABP2 signal intensity was observed in Shank3αβ KO mice compared to wild types (left panel). Significantly higher ZONULIN-1 levels were found in Shank3αβ KO mice (right panel) ( t -test, p = 0.0413). ( F ) The levels of CLAUDIN3 and lipopolysaccharide (LPS) were not significantly different between Shank3αβ KO mice and wild types in gut epithelium. ( G ) Significantly higher ZONULIN-1 levels in Shank3αβ KO mice were confirmed by western blotting using gut epithelium protein lysate ( t -test, p = 0.0434, n = 3 per group). ( H ) Protein lysate from liver tissue from WT and Shank3αβ KO mice ( n = 3 per group) were analyzed for E. coli LPS levels using Western Blotting. The results show significantly higher LPS levels in the liver of Shank3αβ β KO mice ( t -test, p = 0.0452). * p < 0.05, ** p < 0.01.

Article Snippet: Zonulin 1 antibody was purchased from Thermo Fisher Scientific (Invitrogen) (Waltham, MA, USA); Claudin3 antibody from Abcam (Berlin, Germany); FABP2 antibody from Thermo Fisher Scientific (Invitrogen); LPS antibody from Origene (Rockville, MD, USA); and IL6 antibody was purchased from Cell signaling Technologies (Danvers, MA, USA); GFAP antibody was purchased from Sigma Aldrich (St. Louis, MO, USA); Cytokeratin and Vimentin antibody from Abcam.

Techniques: Knock-Out, Staining, MANN-WHITNEY, Immunohistochemistry, Western Blot

Journal: Cellular and Molecular Life Sciences

Article Title: Polysialic acid and Siglec-E orchestrate negative feedback regulation of microglia activation

doi: 10.1007/s00018-020-03601-z

Figure Lengend Snippet: Validation of polySia expression and polysialylated proteins in BV2 microglia. a Immunofluorescence staining of polySia co-localized with the Golgi marker giantin. Cell shape is highlighted by co-staining with the microglia/macrophage marker CD11b. Nuclei were counterstained with DAPI (blue). Scale bar, 20 µm. b Immunoprecipitation (IP) of polysialylated proteins from lysate of 10 7 BV2 cells using polySia-specific mAb 735-conjugated magnetic beads followed by Western blot (WB) detection with polySia-specific antibody (left), or by joint incubation with NRP2- and ESL-1-specific antibodies (right). Where indicated, IP fractions were treated with endosialidase (endo +), to remove polysialic acid. Protein bands were assigned according to the apparent molecular weights of NRP2 and ESL-1 as previously detected in primary and stem cell-derived murine microglia or in mouse brain tissue [ ; see text for details]. c Compared to untreated controls (ctrl), incubation of BV2 cultures with 1-µg/ml LPS for 24 h leads to the loss of polySia signals in almost all cells. Nuclei were counterstained with DAPI (blue). Scale bar, 50 µm. d Loss of polySia-positive cells after LPS treatment, as indicated. Individual values and means of 5 evaluated frames per culture condition, with a minimum of 20 cells each are plotted and significant difference by two-tailed t-test is indicated (*** p < 0.001). e PolySia with DP > 4 but not trisialic acid (DP3) attenuates the LPS-induced production of NO. Nitrite levels in the supernatant of BV2 cells cultured for 24 h in the presence or absence of 30-nM trisialic acid (DP3), 500-ng/ml poly—Sia (approximately 30 nM, see text for details), and/or 1-µg/ml LPS, as indicated. Individual values and means from 3 independent treatments per group are plotted. One-way ANOVA indicated significant differences ( p < 0.0001) and results from Tukey’s post hoc test are shown for comparisons between the LPS-treated groups (* p < 0.05)

Article Snippet: Lipopolysaccharide (LPS) extracted from E. coli serotype O127:B8, 4-chloro- m -cresol (4-C m C), TAK-242, 1,1′‐diheptyl‐4,4′‐bipyridinium dibromide (DHBP), genistein, and biotinyl tyramide were from Merck, Darmstadt, Germany.

Techniques: Biomarker Discovery, Expressing, Immunofluorescence, Staining, Marker, Immunoprecipitation, Magnetic Beads, Western Blot, Incubation, Derivative Assay, Two Tailed Test, Cell Culture

Journal: Cellular and Molecular Life Sciences

Article Title: Polysialic acid and Siglec-E orchestrate negative feedback regulation of microglia activation

doi: 10.1007/s00018-020-03601-z

Figure Lengend Snippet: PolySia staining patterns in BV2 cells treated for 10 or 20 min with 1-µl/ml DMSO ( a ), 50-µM RyR agonist 4-C m C ( b ), 1-µg/ml LPS ( c ), or 1-µg/ml LPS together with 10-µM RyR antagonist DHBP ( d ), and quantitative assessment ( e ), as indicated. Per well, a minimum of 10 cells in 3 randomly selected frames with at least 3 cells each were evaluated and percentages of cells with detectable polySia signals at the cell surface (upper graph) or co-localized with the Golgi marker giantin (lower graph) were calculated. Individual values and means of 3 wells per condition are plotted. For each data set, one-way ANOVA indicated significant differences ( p < 0.0001) and results from Tukey’s post hoc test are shown for comparisons against the DMSO control and for selected group comparisons (** p < 0.01, *** p < 0.001, **** p < 0.0001). 4-C m C and DHBP were added as 1-µl/ml stock solution in DMSO. Giantin and CD11b were co-stained to visualize the Golgi compartment and the cells’ shape, respectively. Nuclei were counterstained with DAPI (blue). Scale bar, 20 µm. See text for a description of treatment effects

Article Snippet: Lipopolysaccharide (LPS) extracted from E. coli serotype O127:B8, 4-chloro- m -cresol (4-C m C), TAK-242, 1,1′‐diheptyl‐4,4′‐bipyridinium dibromide (DHBP), genistein, and biotinyl tyramide were from Merck, Darmstadt, Germany.

Techniques: Staining, Marker, Control

Journal: Cellular and Molecular Life Sciences

Article Title: Polysialic acid and Siglec-E orchestrate negative feedback regulation of microglia activation

doi: 10.1007/s00018-020-03601-z

Figure Lengend Snippet: Detection of polysialylated proteins released by LPS-induced BV2 microglia. a Elution profile of cell culture supernatants collected from 2.5 × 10 7 BV2 cells treated with 10-µg/ml LPS for 24 h and applied to immunoaffinity chromatography with polySia-specific antibody. The increase of conductivity (brown line) denotes the onset of elution with 2-M NaCl. Detection at 214 nm and 280 nm (red and blue line), indicative for the presence of sialic acid and protein, respectively, resulted in peaks during washing (fractions 18–20) and during elution (fractions 22–24). b Western blot detection of polysialylated protein in the cell culture supernatant prior to immunopurification (left panel) and in the pooled fractions 22–24, but not in fractions 18–20 and 26–28 of the chromatogram shown in a (right panel). Specificity of polySia detection in the supernatant was controlled by enzymatic removal of polySia with endosialidase (+ endo). c Elution profile of cell culture supernatants as in a , but collected from 3 × 10 7 BV2 cells during the first 4 h or, after changing the medium, between 4 and 24 h after the onset of LPS treatment, respectively. The left panel shows an elution profile without sample (blank)

Article Snippet: Lipopolysaccharide (LPS) extracted from E. coli serotype O127:B8, 4-chloro- m -cresol (4-C m C), TAK-242, 1,1′‐diheptyl‐4,4′‐bipyridinium dibromide (DHBP), genistein, and biotinyl tyramide were from Merck, Darmstadt, Germany.

Techniques: Cell Culture, Chromatography, Western Blot, Immu-Puri

Journal: Cellular and Molecular Life Sciences

Article Title: Polysialic acid and Siglec-E orchestrate negative feedback regulation of microglia activation

doi: 10.1007/s00018-020-03601-z

Figure Lengend Snippet: LPS-induced changes of Siglec-E in BV2 microglia. a Analysis by quantitative real-time RT-PCR reveals a strong increase of Siglec-E mRNA in cells treated with 1-µg/ml LPS for 24 h. Individual values and means from 3 independent treatments per group are plotted. ** p < 0.01, unpaired t test. b Immunofluorescence detection of Siglec-E (red) and polySia (green). Nuclear counterstain with DAPI (blue). Staining patterns before (control, ctrl) and after treatment with 1-µg/ml LPS for 24 h. Lower panels show 3D reconstructions at higher magnification. c Colocalization of Siglec-E (red) and polySia (green) with the endosomal marker EEA1 (cyan). Staining patterns before (control, ctrl) and after treatment with 1-µg/ml LPS for 10 min. Lower panels show 3D reconstructions of single and merged channels with nuclear counterstain (DAPI, blue) at higher magnification. d – h Detection of Siglec-E (red) and polySia (green) as in b . d Staining patterns after 20 min of LPS treatment in the presence of solvent (1-µl/ml DMSO, left) or 200-µM genistein (right). e , f Staining patterns after incubation for 20 h without or with LPS followed by 10 min with 1-µl/ml DMSO ( e ), 200-µM genistein, or 1-µM TAK-242 ( f ), as indicated. Genistein and TAK-242 were added as 1-µl/ml stock solution in DMSO. g Incubation of LPS-treated cells with DMSO and TAK-242 as in e and f , but this time the cell culture medium was changed to apply DMSO and TAK-242. h Staining patterns after incubation with polySia (10 µg/ml) for 1 min (left) or 20 min (right). The strong Siglec-E signals under control conditions without LPS ( b , c , e , h ) and after genistein treatment ( d , f ) are overexposed to enable a visualization of the weak signals in LPS-treated cells with the same camera setting. Scale bars, 50 µm in b and c (upper panels) and in d – h ; 10 µm in b and c, lower panels. i Quantitative assessment of reduced Siglec-E cell surface staining under the conditions shown in d – h . Based on the densitometric evaluation of signal intensities of 108 cells treated for 10 min with DMSO in the absence of LPS, intensities below 50% of the mean Siglec-E signal intensity under these conditions were considered “reduced”. Per well, a minimum of 20 cells in 3 randomly selected frames with at least 5 cells each were evaluated and the percentage of cells with reduced Siglec-E immunoreactivity was calculated. Individual values and means of 3 wells per condition are plotted. One-way ANOVA indicated significant differences ( p < 0.0001) and results from Tukey’s post hoc test are shown for selected comparisons (**** p < 0.0001)

Article Snippet: Lipopolysaccharide (LPS) extracted from E. coli serotype O127:B8, 4-chloro- m -cresol (4-C m C), TAK-242, 1,1′‐diheptyl‐4,4′‐bipyridinium dibromide (DHBP), genistein, and biotinyl tyramide were from Merck, Darmstadt, Germany.

Techniques: Quantitative RT-PCR, Immunofluorescence, Staining, Control, Marker, Solvent, Incubation, Cell Culture

Journal: Cellular and Molecular Life Sciences

Article Title: Polysialic acid and Siglec-E orchestrate negative feedback regulation of microglia activation

doi: 10.1007/s00018-020-03601-z

Figure Lengend Snippet: Loss of Siglec-E abrogates responsiveness to polySia and enhances LPS-induced activation. a Compared to wildtype BV2 cells ( Siglece +/+ ), the immunoreactivity of Siglec-E (red), but not polySia (green) is abolished by CRISPR/spCas9-mediated knockout of Siglece ( Siglece −/− , clone D19). Nuclear counterstain with DAPI (blue). Scale bar, 50 µm. b Comparable to the effect of preincubation with 60-µM minocycline for 2 h, application of polySia (5 µg/ml) inhibits the LPS-induced NO production of Siglece +/+ but not Siglece −/− BV2 microglia (clone D19). In addition, the LPS-induced NO production of Siglece −/− microglia was significantly higher. Where indicated (LPS +), 1-µg/ml LPS was applied for 24 h. c , d During 24 h of LPS treatment, Siglec-E-negative cells also showed a significantly more pronounced increase of TNF and IL-6 mRNA levels. In b – d , individual values and means from n = 3 independent treatments per group are plotted. Mixed two-way ANOVA indicated significant differences and results from Holms–Sidak post hoc test are shown for selected group comparisons (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001)

Article Snippet: Lipopolysaccharide (LPS) extracted from E. coli serotype O127:B8, 4-chloro- m -cresol (4-C m C), TAK-242, 1,1′‐diheptyl‐4,4′‐bipyridinium dibromide (DHBP), genistein, and biotinyl tyramide were from Merck, Darmstadt, Germany.

Techniques: Activation Assay, CRISPR, Knock-Out