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: Pathogens

Article Title: Distribution and Severity of Placental Lesions Caused by the Chlamydia abortus 1B Vaccine Strain in Vaccinated Ewes

doi: 10.3390/pathogens10050543

Figure Lengend Snippet: Histopathological changes and IHC labelling in the placentas of sheep infected with C. abortus vaccine-type strains. ( A , C , E ) Placenta vt-P1: showing ( A , C ) necrosuppurative placentitis with a large numbers of leucocytes in the chorionic epithelium of the cotyledon attached to superficial amorphous necrotic material stained by HE technique; ( E ) chlamydial organisms labelled by IHC using anti-LPS mAb 13/4 and counterstained with haematoxylin; ( B , D , F ) Placenta vt-P2: showing ( B , D ) necrosuppurative placentitis and extensive compact layer of leucocytes in the trophoblast layer and basement membrane stained by HE technique and ( F ) chlamydial organisms labelled by IHC using anti-LPS mAb 13/4 and counterstained with haematoxylin. Black outlined areas shown in top images indicates areas expanded in images located immediately below. Scale bars are as indicated.

Article Snippet: IHC was performed in the same placental areas detailed in and in using a genus-specific mouse monoclonal antibody raised against the lipopolysaccharide (LPS) of C. abortus strain S26/3 (mAb 13/4, Santa Cruz Biotechnology, Heidelberg, Germany), as previously described [ ], but using goat anti-mouse IgG conjugate (EnvisionTM + System HRP labelled polymer, Dako, Ely, UK) with aminoethyl carbazole alcohol soluble chromogen (AEC, Vector Laboratories, Peterborough, UK) enzyme substrate for the labelling.

Techniques: Infection, Staining, Membrane

Journal: Pathogens

Article Title: Distribution and Severity of Placental Lesions Caused by the Chlamydia abortus 1B Vaccine Strain in Vaccinated Ewes

doi: 10.3390/pathogens10050543

Figure Lengend Snippet: PCA biplots for trophoblast and mesenchyme layers in cotyledonary ( A : CT, C : CM) and intercotyledonary ( B : ICT, D : ICM) areas. The parameters analysed are necrosis (N), infiltration of polymorphonuclear leukocytes (PMNI), infiltration of mononuclear cells (MI), and labelling of chlamydial LPS by IHC (TMIHC). Note that the four biplots show very comparable patterns in terms of both, configuration of the treatment group points and associations between pathology parameters. The percentage of explained data variability by each PC axis is indicated in parenthesis.

Article Snippet: IHC was performed in the same placental areas detailed in and in using a genus-specific mouse monoclonal antibody raised against the lipopolysaccharide (LPS) of C. abortus strain S26/3 (mAb 13/4, Santa Cruz Biotechnology, Heidelberg, Germany), as previously described [ ], but using goat anti-mouse IgG conjugate (EnvisionTM + System HRP labelled polymer, Dako, Ely, UK) with aminoethyl carbazole alcohol soluble chromogen (AEC, Vector Laboratories, Peterborough, UK) enzyme substrate for the labelling.

Techniques:

Journal: Pathogens

Article Title: Distribution and Severity of Placental Lesions Caused by the Chlamydia abortus 1B Vaccine Strain in Vaccinated Ewes

doi: 10.3390/pathogens10050543

Figure Lengend Snippet: PCA biplot for blood vessels in cotyledonary ( A : CT, C : CM) and intercotyledonary ( B : ICT, D : ICM) areas. The parameters analysed are mural necrosis (VN), vasculitis-infiltration of polymorphonuclear leukocytes (VPMNI), vasculitis-infiltration of mononuclear cells (VMI), vascular endothelial activation (VEA), vascular thrombosis (VT) and labelling by IHC for C. abortus LPS (VIHC). Note that the four biplots show very comparable patterns in terms of both configuration of the treatment group points and associations between pathology parameters. The percentage of explained data variability by each PC axis is indicated in parenthesis.

Article Snippet: IHC was performed in the same placental areas detailed in and in using a genus-specific mouse monoclonal antibody raised against the lipopolysaccharide (LPS) of C. abortus strain S26/3 (mAb 13/4, Santa Cruz Biotechnology, Heidelberg, Germany), as previously described [ ], but using goat anti-mouse IgG conjugate (EnvisionTM + System HRP labelled polymer, Dako, Ely, UK) with aminoethyl carbazole alcohol soluble chromogen (AEC, Vector Laboratories, Peterborough, UK) enzyme substrate for the labelling.

Techniques: Activation Assay

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO promotes lysosomal degradation of SDC4-CTF. A , Western blot of SDC1/4 CTF and full-length in SW480 cells treated with AGO (indicated doses, 24 h; n = 3). B , Western blot in SW480 cells treated with 20 μM AGO for 0 to 24 h. C and D , immunofluorescence detection and quantification of SDC4 in SW480 cells treated with 20 μM AGO (24 h; n = 6). E , surface SDC4 measured by FACS using FITC-anti-SDC4. F , Western blot of SDC1/4 CTF and full-length in HCT116 cells treated with AGO (indicated doses, 24 h). G , Western blot of SDC4-CTF in multiple colorectal cancer cells treated with 20 μM AGO (24 h). H , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with cycloheximide (50 μg/ml) along with/without 20 μM AGO for indicated times. I , RT-qPCR of indicated genes in SW480 cells treated with 20 μM AGO (24 h; n = 4). J , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with 20 μM AGO, 2.5 μM GM6001, 10 μM TMI-1, or combinations (24 h). K , Western blot in SW480 cells treated with 20 μM AGO, 5 μM BAY11 to 7082, 10 nM PMA, 1 μg/ml LPS, or 20 μg/ml TNFα (24 h). L and M , live-cell imaging and analysis of SDC4-GFP intensity in 293T cells co-expressing DsRed-Rab5A and SDC4-GFP, with 20 μM AGO treatment (n = 4). O , Western blot of lysosome markers and SDC1/4-CTF in SW480 cells treated with 20 μM AGO, 250 nM BAF1, or combination (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns). AGO, andrographolide; SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; FACS, fluorescence-activated cell sorting; BAF1, Bafilomycin A1.

Article Snippet: Andrographolide (HY-N0191), MG132 (HY-13259), Bortezomib (HY-10227), Carfilzomib (HY-10455), Ixazomib (HY-10453), DAPT (HY-13027), GM6001 (HY-15768), TMI-1 (HY-101448), LY294002 (HY-10108), Rapamycin (HY-10219), Bafilomycin A1 (HY-100558), cycloheximide (HY-12320), BAY 11 to 7082 (HY-13453), PMA (HY-18739), LPS (HY-D1056), and TNFα (HY- P70426 ) were purchased from MedChemExpress.

Techniques: Western Blot, Immunofluorescence, Quantitative RT-PCR, Live Cell Imaging, Expressing, Two Tailed Test, Fluorescence, FACS

Journal: bioRxiv

Article Title: The Chlamydia trachomatis secreted effector protein CT181 binds to Mcl-1 to prolong neutrophil survival

doi: 10.1101/2025.03.16.643443

Figure Lengend Snippet: ( A ) A2EN or HeLa cells were infected at a MOI of 2.5 with wild-type, tmeA-lx , CT181:: bla , or CT144:: bla . At 48h, cells were lysed and replated on fresh HeLa cell monolayers and infectious forming units were quantified by immunofluorescence microscopy. ( B ) A2EN or HeLa cells were infected at a MOI of 2.5 for 60 min with wild-type, tmeA-lx , CT181:: bla , or CT144:: bla . The number of internal bacteria was determine using differential immunostaining. ( C) To measure inclusion size, A2EN cells were infected at an MOI of 2.5 for 24 h. Bacteria were stained with anti-LPS (green), the inclusion membrane was stained with an anti-IncE antibody (red), and DNA was stained with DAPI (blue). Inclusion diameter was measured in ImageJ. ( A-C ) Data are representative of three independent experiments. Statistical significance was determined using One-Way ANOVA with Dunnett’s multiple comparison post-test comparing the mutants to wild-type. ****P<0.0001, ***P<0.001, **P<0.01, *P<0.05.

Article Snippet: LPS antibody (Novus Cat#NBP1-28820) for enumeration of inclusions by immunofluorescence microscopy.

Techniques: Infection, Immunofluorescence, Microscopy, Bacteria, Immunostaining, Staining, Membrane, Comparison

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