Journal: Brain, behavior, and immunity

Article Title: Alcohol Exposure after Mild Focal Traumatic Brain Injury Impairs Neurological Recovery and Exacerbates Localized Neuroinflammation

doi: 10.1016/j.bbi.2014.11.006

Figure Lengend Snippet: Diagram of coronal rat brain section showing the site of injury and photographed region (4 A). Representative immunofluorescence images (10 ×) of ipsilateral cortex GFAP, ED-1, and HMGB1 immunoreactivity (green) combined with DAPI (blue) at 14 days post-TBI (10 days post-alcohol exposure) (4 B).

Article Snippet: Anti-HMGB1 Rabbit polyclonal (Abcam, Cambridge, UK) , 1:100 , Donkey anti-Rabbit Alexa Fluor 488 (Invitrogen, Carlsbad, CA) , 1:200.

Techniques: Immunofluorescence

Journal: Brain, behavior, and immunity

Article Title: Alcohol Exposure after Mild Focal Traumatic Brain Injury Impairs Neurological Recovery and Exacerbates Localized Neuroinflammation

doi: 10.1016/j.bbi.2014.11.006

Figure Lengend Snippet: Quantification of ipsilateral cortex GFAP, ED-1, and HMGB1 immunoreactivity at 14 days post-TBI (10 days post-alcohol exposure). Images are quantified as % area of positive staining in 1.035 mm2 (10×). Values are shown as means ± SEM. For GFAP and ED-1, Sham/Air n = 4, Sham/Alcohol n = 4, TBI/Air n = 4, TBI/Alcohol n = 6. For HMGB1, Sham/Air n = 4, Sham/Alcohol n = 5, TBI/Air n = 5, TBI/Alcohol n = 7 (each n represents the average of 3 images taken from each animal) * p < 0.05 of the TBI groups vs. time-matched sham controls; $ p < 0.05 of the TBI/Alcohol group vs. TBI/Air group, by two-way ANOVA (5 A, 5 C, and 5 E).

Article Snippet: Anti-HMGB1 Rabbit polyclonal (Abcam, Cambridge, UK) , 1:100 , Donkey anti-Rabbit Alexa Fluor 488 (Invitrogen, Carlsbad, CA) , 1:200.

Techniques: Staining

Journal: Brain, behavior, and immunity

Article Title: Alcohol Exposure after Mild Focal Traumatic Brain Injury Impairs Neurological Recovery and Exacerbates Localized Neuroinflammation

doi: 10.1016/j.bbi.2014.11.006

Figure Lengend Snippet: Primary and Secondary Antibodies.

Article Snippet: Anti-HMGB1 Rabbit polyclonal (Abcam, Cambridge, UK) , 1:100 , Donkey anti-Rabbit Alexa Fluor 488 (Invitrogen, Carlsbad, CA) , 1:200.

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Role of Myeloperoxidase Oxidants in the Modulation of Cellular Lysosomal Enzyme Function: A Contributing Factor to Macrophage Dysfunction in Atherosclerosis?

doi: 10.1371/journal.pone.0168844

Figure Lengend Snippet: (A) Cathepsin B and (B) cathepsin L activity in J774A.1 cells (1 × 10 6 cells mL -1 ) was determined after incubation with HOSCN (80–160 μM, white bars) or HOCl (80–160 μM, black bars) for 15 min at 22°C. (C) Cathepsin B and (D) cathepsin L activity in J774A.1 cell lysates (1 × 10 6 cells mL -1 ) after incubation with HOSCN (5–20 μM) for 15 min, followed by further incubation in the absence (white bars) or presence (black bars) of DTT (100 μM) for 15 min. Results are expressed as a percentage of the PBS-treated control cells. * and # represent a significant (p < 0.05) change in cathepsin B/L activity compared with control lysates or the presence / absence of DTT, respectively.

Article Snippet: And incubation with either anti-cathepsin B goat polyclonal (Santa Cruz Biotechnology, Dallas, Texas, USA), anti-cathepsin L mouse monoclonal (Abcam, Cambridge, UK), anti-LAMP-1 rabbit polyclonal (Abcam) or anti-β-actin mouse monoclonal (Santa Cruz) primary antibodies (1/1000 dilution).

Techniques: Activity Assay, Incubation

Journal: PLoS ONE

Article Title: Role of Myeloperoxidase Oxidants in the Modulation of Cellular Lysosomal Enzyme Function: A Contributing Factor to Macrophage Dysfunction in Atherosclerosis?

doi: 10.1371/journal.pone.0168844

Figure Lengend Snippet: LDL (1 mg protein mL -1 ) was exposed to 0–500 μM HOSCN (A and B) or HOCl (C and D) for 30 min (white) and 24 h (black) at 22°C and 37°C respectively, prior to addition of each modified LDL (0.1 mg protein mL -1 ) to J774A.1 lysates for 15 min at 22°C, followed by determination of cathepsin B (A, C) or cathepsin L (B, D) activity, which is expressed relative to the no LDL control. Data are means ± SEM for at least 3 independent experiments, with multiple LDL donors. * and # represent a significant (p < 0.05) decrease in cathepsin B or L activity compared to cells exposed to the incubation control LDL or no LDL. “a” represents a significant (p < 0.05) difference between 30 min and 24 h modified LDL.

Article Snippet: And incubation with either anti-cathepsin B goat polyclonal (Santa Cruz Biotechnology, Dallas, Texas, USA), anti-cathepsin L mouse monoclonal (Abcam, Cambridge, UK), anti-LAMP-1 rabbit polyclonal (Abcam) or anti-β-actin mouse monoclonal (Santa Cruz) primary antibodies (1/1000 dilution).

Techniques: Modification, Activity Assay, Incubation

Journal: PLoS ONE

Article Title: Role of Myeloperoxidase Oxidants in the Modulation of Cellular Lysosomal Enzyme Function: A Contributing Factor to Macrophage Dysfunction in Atherosclerosis?

doi: 10.1371/journal.pone.0168844

Figure Lengend Snippet: LDL (1 mg protein mL -1 ) was exposed to 0–5000 μM KOCN for 24 h at 37°C, prior to addition of each modified LDL (0.1 mg protein mL -1 ) to J774A.1 lysates for 15 min at 22°C, followed by determination of cathepsin B (white bars) or cathepsin L (black bars) activity, which is expressed relative to the no LDL control. * and # represent a significant decrease (p < 0.05) in cathepsin B or L activity compared with cells exposed to the incubation control LDL or no LDL respectively by 1-way ANOVA with Tukey’s post-hoc testing.

Article Snippet: And incubation with either anti-cathepsin B goat polyclonal (Santa Cruz Biotechnology, Dallas, Texas, USA), anti-cathepsin L mouse monoclonal (Abcam, Cambridge, UK), anti-LAMP-1 rabbit polyclonal (Abcam) or anti-β-actin mouse monoclonal (Santa Cruz) primary antibodies (1/1000 dilution).

Techniques: Modification, Activity Assay, Incubation

Journal: PLoS ONE

Article Title: Role of Myeloperoxidase Oxidants in the Modulation of Cellular Lysosomal Enzyme Function: A Contributing Factor to Macrophage Dysfunction in Atherosclerosis?

doi: 10.1371/journal.pone.0168844

Figure Lengend Snippet: LDL (1 mg protein mL -1 ) was exposed to 0–500 μM HOSCN (A and B) or HOCl (C and D) for 24 h at 37°C respectively, prior to addition of each modified LDL (0.1 mg protein mL -1 ) to J774A.1 cells for 4 (white bars) or 24 h (black bars), and determination of cathepsin B (A, C) or cathepsin L (B, D) activity, which is expressed relative to the no LDL control. * and # represent a significant decrease (p < 0.05) in cathepsin B or L activity compared with cells exposed to the incubation control LDL or no LDL. “a” represents a significant (p < 0.05) difference between cathepsin activity between cells incubated with LDL for 4 or 24 h.

Article Snippet: And incubation with either anti-cathepsin B goat polyclonal (Santa Cruz Biotechnology, Dallas, Texas, USA), anti-cathepsin L mouse monoclonal (Abcam, Cambridge, UK), anti-LAMP-1 rabbit polyclonal (Abcam) or anti-β-actin mouse monoclonal (Santa Cruz) primary antibodies (1/1000 dilution).

Techniques: Modification, Activity Assay, Incubation

Journal: PLoS ONE

Article Title: Role of Myeloperoxidase Oxidants in the Modulation of Cellular Lysosomal Enzyme Function: A Contributing Factor to Macrophage Dysfunction in Atherosclerosis?

doi: 10.1371/journal.pone.0168844

Figure Lengend Snippet: LDL (1 mg protein mL -1 ) was exposed to 0–500 μM HOSCN (A) or HOCl (B) for 24 h at 37°C, respectively, prior to addition of each modified LDL (0.1 mg protein mL -1 ) to J774A.1 cells, followed by determination of cathepsin B (white) and L (black) protein expression. Cathepsin levels were normalised to β-actin levels, and then calculated as the fold change from the no LDL condition. Representative blots of the cathepsin B band at 25 kDa, the cathepsin L band at 25 kDa, or the β-actin band at 43 kDa, are displayed, of n = 3–4 separate experiments. There was no significant effect of oxidant treatment as determined by 1-way ANOVA on either cathepsin protein levels.

Article Snippet: And incubation with either anti-cathepsin B goat polyclonal (Santa Cruz Biotechnology, Dallas, Texas, USA), anti-cathepsin L mouse monoclonal (Abcam, Cambridge, UK), anti-LAMP-1 rabbit polyclonal (Abcam) or anti-β-actin mouse monoclonal (Santa Cruz) primary antibodies (1/1000 dilution).

Techniques: Modification, Expressing