Journal: Experimental neurology

Article Title: Impaired Social Behaviors and Minimized Oxytocin Signaling of the Adult Mice Deficient in the N-Methyl-D-Aspartate Receptor GluN3A Subunit

doi: 10.1016/j.expneurol.2018.02.015

Figure Lengend Snippet: Western blot analysis was performed on brain tissues from the hippocampus and thalamus areas of WT and GluN3A KO mice. A. Western blot images and quantification of the band intensity compared to the basal level. The protein level of oxytocin receptor, serotonin receptors 5-HTR2A, 5-HTR1B, TNFR1 and CD73 showed no statistically difference between WT and KO mice. The level of CD38, however, was significantly higher in the KO hippocampus. N=6, Student t test; * p<0.05, F=1.299. B. The above assay was repeated in the thalamus. All measured protein expressions were similar between WT and KO mice, expect the 5-HTR2A level was significantly lower in this brain region. N=6; Student t test; * p<0.05, F=5.650.

Article Snippet: The primary antibodies used and the dilutions for each were mouse anti-serotonin receptor (5-HTR; BD Bioscience, San Jose, CA, USA) at 1:2,500, rabbit anti-serotonin transporter (BD Bioscience) at 1:1,000, rabbit anti-oxytocin antibody (Fitzgerald Industries International, North Acton, MA, USA) at 1:1000, goat anti-oxytocin receptor antibody (Abcam, Cambridge, MA, USA) at 1:2,000, goat anti-CD73 antibody (Santa Cruz Biotech, Inc., Dallas, TX, USA) at 1:2,000, rabbit anti-BDNF antibody (Santa Cruz Biotech) at 1:2,000, mouse anti-TNFR1 antibody (Santa Cruz Biotech) at 1:2,000, rabbit anti-GluN3A antibody (Abcam) at 1:2,000, and mouse anti-actin (Sigma-Aldrich).

Techniques: Western Blot

Journal: Experimental neurology

Article Title: Impaired Social Behaviors and Minimized Oxytocin Signaling of the Adult Mice Deficient in the N-Methyl-D-Aspartate Receptor GluN3A Subunit

doi: 10.1016/j.expneurol.2018.02.015

Figure Lengend Snippet: We investigated the possibility that the deletion of GluN3A could altered social behavior-associated genes such as serotonin receptor (5-HTR), serotonin transporter (5-HTT), BDNF, CD73, and TNFR1. A. Western blotting of serotonin receptor, serotonin transporter, BDNF, CD73, and TNFR1 in the prefrontal cortex from WT and GluN3A KO mice. B-F. Quantification of the optic density (normalized to β-actin control) of 5-HTR, 5-HTT, BDNF, CD73, and TNFR1 in the prefrontal cortex. The expression of these genes was also unchanged in GluN3A KO mice. N=6 per group.

Article Snippet: The primary antibodies used and the dilutions for each were mouse anti-serotonin receptor (5-HTR; BD Bioscience, San Jose, CA, USA) at 1:2,500, rabbit anti-serotonin transporter (BD Bioscience) at 1:1,000, rabbit anti-oxytocin antibody (Fitzgerald Industries International, North Acton, MA, USA) at 1:1000, goat anti-oxytocin receptor antibody (Abcam, Cambridge, MA, USA) at 1:2,000, goat anti-CD73 antibody (Santa Cruz Biotech, Inc., Dallas, TX, USA) at 1:2,000, rabbit anti-BDNF antibody (Santa Cruz Biotech) at 1:2,000, mouse anti-TNFR1 antibody (Santa Cruz Biotech) at 1:2,000, rabbit anti-GluN3A antibody (Abcam) at 1:2,000, and mouse anti-actin (Sigma-Aldrich).

Techniques: Western Blot, Expressing

Journal: Toxicology

Article Title: Differential Fmo3 Gene Expression in Various Liver Injury Models Involving Hepatic Oxidative Stress in Mice

doi: 10.1016/j.tox.2014.08.013

Figure Lengend Snippet: Livers were collected from mice (n=6) sacrificed at respective time-points (2,4,8,12,24 & 48h for ANIT; 24 & 48h for CCl4; 6 & 24h for AlOH; and 10 d for BDL). RNA was isolated and cDNA was made using a commercial MMLV-RT kit. The cDNA samples were analyzed by quantitative RT-PCR using Fmo3 mouse-specific primers. Gene expression was normalized to the housekeeping gene β-actin. Fmo3 mRNA expression is presented as mean Fold Change ± SE. One-way ANOVA, t-test or two-way ANOVA was performed, appropriately, followed by the Dunnett's posttest for One-way ANOVA and the Bonferroni posttest for two-way ANOVA. Asterisks (*) represent a statistical difference (p < 0.05) between vehicle-treated and hepatotoxicant-treated or BDL group.

Article Snippet: Membranes were blocked with 5 % non-fat powdered milk in tris buffered saline containing 0.05% tween-20(TBS-T) for 8 h. A rabbit anti-mouse Fmo3 primary antibody (GenScript USA Inc., NJ) (1:5000) was used to detect Fmo3 with β-actin as a loading control.

Techniques: Isolation, Quantitative RT-PCR, Expressing

Journal: Toxicology

Article Title: Differential Fmo3 Gene Expression in Various Liver Injury Models Involving Hepatic Oxidative Stress in Mice

doi: 10.1016/j.tox.2014.08.013

Figure Lengend Snippet: Western immunoblots for Fmo3 were performed using liver microsomes from control and hepatotoxicant-treated or BDL mice. A custom-made rabbit anti-mouse Fmo3 primary antibody, described in Materials and Methods was used to detect Fmo3. Fmo3 protein levels were normalized to β-actin loading control. Microsomal proteins isolated from naïve female mouse liver were used as a positive control (indicated by “+” sign). The data are presented as blots and as mean Fmo3 protein expression (Fold Change) ± SE. One-way ANOVA, t-test or two-way ANOVA was performed, appropriately, followed by the Dunnett's posttest for One-way ANOVA and the Bonferroni posttest for two-way ANOVA. Asterisks (*) represent a statistical difference (p < 0.05) between vehicle-treated and hepatotoxicant-treated or BDL group.

Article Snippet: Membranes were blocked with 5 % non-fat powdered milk in tris buffered saline containing 0.05% tween-20(TBS-T) for 8 h. A rabbit anti-mouse Fmo3 primary antibody (GenScript USA Inc., NJ) (1:5000) was used to detect Fmo3 with β-actin as a loading control.

Techniques: Western Blot, Isolation, Positive Control, Expressing

Journal: Toxicology

Article Title: Differential Fmo3 Gene Expression in Various Liver Injury Models Involving Hepatic Oxidative Stress in Mice

doi: 10.1016/j.tox.2014.08.013

Figure Lengend Snippet: Plasma and livers were collected from mice 72 h following APAP (400 mg/kg) or vehicle treatment. (A) The data are presented as mean plasma ALT (IU/L) ± SE. (B) RNA was isolated from livers andcDNA samples were analyzed by quantitative RT-PCR using Fmo3 mouse-specific primers. Gene expression was normalized to the housekeeping gene β-actin. Fmo3 mRNA expression are presented as mean Fold Change ± SE. Oneway ANOVA was performed followed by the Dunnett's post-test. Asterisks (*) represent a statistical difference (p < 0.05) between vehicle-treated group and APAP-treated group and hash (#) represent a statistical difference (p<0.05) compared with APAP-treated wild-type mice.

Article Snippet: Membranes were blocked with 5 % non-fat powdered milk in tris buffered saline containing 0.05% tween-20(TBS-T) for 8 h. A rabbit anti-mouse Fmo3 primary antibody (GenScript USA Inc., NJ) (1:5000) was used to detect Fmo3 with β-actin as a loading control.

Techniques: Isolation, Quantitative RT-PCR, Expressing

Journal: Toxicology

Article Title: Differential Fmo3 Gene Expression in Various Liver Injury Models Involving Hepatic Oxidative Stress in Mice

doi: 10.1016/j.tox.2014.08.013

Figure Lengend Snippet: After overnight fasting, groups of wild-type and Nrf2 knockout mice received a single dose of 400 mg/kg APAP or vehicle. Livers were collected 72 h following APAP or vehicle treatments. Western blot for Fmo3 was performed using liver microsomes from control and APAP-treated mice. Equal protein loading (10 μg protein/lane) was confirmed by detection of β-actin. Microsomal proteins isolated from naïve female mouse liver were used as a positive control indicated by “+” sign. The data are presented as blots and as mean Fmo3 protein expression (Fold Change) ± SE (A). FMO activity was measured in liver microsomes from control and APAP-treated mice using methimazole as substrate. Data are presented as mean Specific Activity (μM/min/mg) ± SE (B). Asterisks (*) represent a statistical difference (p < 0.05) between vehicle-treated group and APAP-treated group and hash (#) represent a statistical difference (p<0.05) compared with APAP-treated wild-type mice.

Article Snippet: Membranes were blocked with 5 % non-fat powdered milk in tris buffered saline containing 0.05% tween-20(TBS-T) for 8 h. A rabbit anti-mouse Fmo3 primary antibody (GenScript USA Inc., NJ) (1:5000) was used to detect Fmo3 with β-actin as a loading control.

Techniques: Knock-Out, Western Blot, Isolation, Positive Control, Expressing, Activity Assay