Journal: Scientific Reports

Article Title: Alpha-1 microglobulin as a potential therapeutic candidate for treatment of hypertension and oxidative stress in the STOX1 preeclampsia mouse model

doi: 10.1038/s41598-019-44639-9

Figure Lengend Snippet: Human rA1M protects kidney function but no effect on cardiac hypertrophy. ( a ) Albumin/Creatinine ratio (ACR) analysis of urine from pregnant females showed an increase of ACR at late-gestation in the preeclamptic females, which was absent in the rA1M-treated group. Shown is the mean ACR normalised to Ctrl-buff values for each gestation period, and n = number of females analysed. ( b ) Gene expression levels for HO-1 , CAT and SOD2 , normalised to the HPRT -gene levels in kidney from PE-buff and PE-A1M females, demonstrating significant reduction of HO-1 and CAT expression (*p = 0.04; **p = 0.04) after rA1M-treatment. The line represents the median and n = number of females analysed. ( c ) Preeclamptic females showed increased heart weight compared to Ctrl-buff at 17.5 dpc (*p = 0.002), which could not be alleviated by rA1M treatment (PE-A1M vs Ctrl-buff; **p = 0.008). Control groups showed similar heart weight as non-pregnant females. The line represents the median and n = number of females analysed.

Article Snippet: Real-time PCR was performed for HO-1 , CAT , SOD2 , with hypoxanthine phosphoribosyltransferase ( HPRT) as endogenous control, using TaqMan Gene Expression Assays specific for mouse ( HO-1 – Mm00516005_m1, CAT – Mm00437992_m1, SOD2 – Mm01313000_m1, HPRT – Mm01545399_m1, Life Technologies).

Techniques: Gene Expression, Expressing, Control

Journal: Cell reports

Article Title: Gut Microbiota Composition Modulates the Magnitude and Quality of Germinal Centers during Plasmodium Infections

doi: 10.1016/j.celrep.2020.108503

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Peroxidase Affini Pure Goat Anti-Mouse IgM , Jackson ImmunoResearch , Cat# 115–035-020; RRID:AB_2338502.

Techniques: Blocking Assay, Recombinant, Staining, Bradford Protein Assay, Sequencing, Generated, Software, Modification, Irradiation

Journal: eLife

Article Title: Plural molecular and cellular mechanisms of pore domain KCNQ2 encephalopathy

doi: 10.7554/eLife.91204

Figure Lengend Snippet:

Article Snippet: Antibody , Peroxidase IgG Fraction Monoclonal Mouse Anti-Rabbit IgG, light chain specific , Jackson ImmunoResearch , Code: 211-032-171 Lot #: 97224 RRID: AB_2339149 , WB (1:5000).

Techniques: Transfection, Construct, Mutagenesis, Recombinant, Expressing, Plasmid Preparation, Gene Expression, Real-time Polymerase Chain Reaction, Sequencing, Western Blot, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Incubation, Software

Journal: EMBO Reports

Article Title: Promyelocytic leukemia protein targets MK2 to promote cytotoxicity

doi: 10.15252/embr.202052254

Figure Lengend Snippet:

Article Snippet: Peroxidase IgG Fraction Monoclonal Mouse Anti‐Goat IgG, light chain specific , Jackson ImmunoResearch , 205‐032‐176, RRID:AB_2339056 , WB 1:20,000.

Techniques: Generated, Transgenic Assay, Recombinant, Plasmid Preparation, Blocking Assay, Transfection, Antibody Labeling, Software, Cell Viability Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Microglial SLC25A28 Knockout Mitigates Spinal Cord Injury in Mice by Inhibiting Heme Synthesis and Subsequent NOX2 Activation

doi: 10.1111/cns.70638

Figure Lengend Snippet: SLC25A28 regulates microglial activation via NOX2‐dependent ROS pathway. (a) CYBB mRNA levels in SLC25A28‐KO versus control microglia (qPCR, n = 3). (b) NOX2 protein expression in microglia (Western blot, n = 3). (c) Intracellular ROS levels in microglia assessed by DCFH‐DA staining (scale bar = 5 μm, n = 5). (d) SLC25A28 mRNA expression in overexpression (OE) versus control BV2 cells (qPCR, n = 3). (e) SLC25A28 protein levels in OE and control cells (Western blot, n = 3). (f) Microglial morphology with/without GSK2795039 (10 μM) treatment; cell body area was quantified (scale bar = 20 μm, n = 3). (g) Mitochondrial iron visualization using Mito‐FerroGreen staining (scale bar = 5 μm). (h) Quantification of mitochondrial iron content by ferrozine assay ( n = 3). (i) CYBB mRNA expression in OE cells with or without GSK2795039 treatment (qPCR, n = 3). (j) NOX2 protein expression in OE cells with or without GSK2795039 treatment (Western blot, n = 3). (k) ROS levels in OE cells with or without GSK2795039 treatment, measured by DCFH‐DA staining (scale bar = 20 μm, n = 6). (l) Expression of inflammatory markers (iNOS, total caspase‐1, and cleaved caspase‐1) in OE cells with or without GSK2795039 treatment (Western blot, n = 3). (m) ELISA quantification of IL‐1β and TNF‐α in OE cells with or without GSK2795039 treatment ( n = 3). Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Specifically, the NOX2 inhibitor GSK2795039 (10 μM, MedChemExpress, HY‐18950, Monmouth Junction, NJ, USA) was added 1 h prior to LPS stimulation in SLC25A28‐overexpressing BV2 cells to suppress NOX2 activity.

Techniques: Activation Assay, Control, Expressing, Western Blot, Staining, Over Expression, Ferrozine Assay, Enzyme-linked Immunosorbent Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Microglial SLC25A28 Knockout Mitigates Spinal Cord Injury in Mice by Inhibiting Heme Synthesis and Subsequent NOX2 Activation

doi: 10.1111/cns.70638

Figure Lengend Snippet: SLC25A28 overexpression promotes NOX2 activation through enhanced heme biosynthesis. (a) ALAS1 mRNA levels in SLC25A28 overexpression (A28 OE) versus control cells (qPCR, n = 3). (b) ALAS1 protein expression in A28 OE and control cells (Western blot, n = 3). (c) Ferrochelatase (FECH) mRNA levels in A28 OE and control cells (qPCR, n = 3). (d) Ferrochelatase protein expression in A28 OE and control cells (Western blot, n = 3). (e) Cellular heme content in A28 OE cells (Oxalate assay, n = 6). (f) Microglial morphology in A28 OE cells with or without succinylacetone (SA) treatment (scale bar = 20 μm, n = 3). (g) NOX2 mRNA levels in A28 OE cells with or without SA treatment (qPCR, n = 3). (h) NOX2 protein expression in A28 OE cells with or without SA treatment (Western blot, n = 3). (i) ROS production in A28 OE cells with or without SA treatment (DCFH‐DA staining, scale bar = 5 μm, n = 6). (j) Inflammatory marker expression in A28 OE cells with or without SA treatment (iNOS, total caspase‐1, and cleaved caspase‐1; Western blot, n = 3). (k) ELISA quantification of IL‐1β and TNF‐α levels in A28 OE cells with or without SA treatment ( n = 3). Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Specifically, the NOX2 inhibitor GSK2795039 (10 μM, MedChemExpress, HY‐18950, Monmouth Junction, NJ, USA) was added 1 h prior to LPS stimulation in SLC25A28‐overexpressing BV2 cells to suppress NOX2 activity.

Techniques: Over Expression, Activation Assay, Control, Expressing, Western Blot, Staining, Marker, Enzyme-linked Immunosorbent Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Microglial SLC25A28 Knockout Mitigates Spinal Cord Injury in Mice by Inhibiting Heme Synthesis and Subsequent NOX2 Activation

doi: 10.1111/cns.70638

Figure Lengend Snippet: SLC25A28 knockout attenuates neuroinflammation through heme‐dependent NOX2 activation. (a) ALAS1 mRNA levels in A28‐KO versus control primary microglia (qPCR, n = 3). (b) ALAS1 protein expression in A28‐KO and control microglia (Western blot, n = 3). (c) FECH mRNA levels in A28‐KO versus control microglia (qPCR, n = 3). (d) FECH protein expression in A28‐KO and control microglia (Western blot, n = 3). (e) Cellular heme content in A28‐KO cells (oxalate assay, n = 6). (f) Microglial morphology in A28‐KO cells with or without 5‐aminolevulinic acid (5‐ALA) treatment (scale bar = 20 μm, n = 3). (g) NOX2 mRNA levels in A28‐KO cells with or without 5‐ALA treatment (qPCR, n = 3). (h) NOX2 protein levels in A28‐KO cells with or without 5‐ALA treatment (Western blot, n = 3). (i) ROS production in A28‐KO cells with or without 5‐ALA treatment (DCFH‐DA staining, scale bar = 5 μm, n = 5). (j) Inflammatory marker expression in A28‐KO cells with or without 5‐ALA treatment (iNOS, total caspase‐1, and cleaved caspase‐1; Western blot, n = 3). (k) ELISA quantification of IL‐1β and TNF‐α secretion in A28‐KO cells with or without 5‐ALA treatment ( n = 3). Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Specifically, the NOX2 inhibitor GSK2795039 (10 μM, MedChemExpress, HY‐18950, Monmouth Junction, NJ, USA) was added 1 h prior to LPS stimulation in SLC25A28‐overexpressing BV2 cells to suppress NOX2 activity.

Techniques: Knock-Out, Activation Assay, Control, Expressing, Western Blot, Staining, Marker, Enzyme-linked Immunosorbent Assay