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

Journal: Cell Communication and Signaling : CCS

Article Title: Presenilin1 exerts antiproliferative effects by repressing the Wnt/β-catenin pathway in glioblastoma

doi: 10.1186/s12964-019-0501-9

Figure Lengend Snippet: a Representative IHC staining of Presenilin1 protein in low grade glioma tissues (LGG, left panels) and high grade glioma tissues (HGG, right panels). Scale bars = 50 um and 20 um respectively. b Statistical analysis of the relative expression levels of Presenilin1 in LGG and HGG, * p < 0.05. c Kaplan-Meier analysis for all grade glioma patients, patients in the high Presenilin1 group ( n = 122) and in the low Presenilin1 group ( n = 89) ( P = 0.0000, log-rank test). d Kaplan-Meier analysis for glioma patients. Patients in the high Presenilin1 group ( n = 28) and in the low Presenilin1 group ( n = 91) ( P = 0.0257, log-rank test). e Kaplan-Meier analysis for all grade glioma patients. Patients in the high Presenilin1 group ( n = 180) and in the low Presenilin1 group ( n = 92) ( P = 0.0019, log-rank test). f U87 cells were transfected with lentivirus contain sh-RNA target Presenilin1(sh-PS1) or full-length gene of Presenilin1(Lv-PS1).Sh-NC and Lv-NC were used as control, then Western-blot to detect the down- and over-expression effect of lentivirus in U87 cells, * p < 0.05,** p < 0.01

Article Snippet: Rabbit anti-Presenilin1 monoclonal antibody was purchased from Proteintech (Wuhan, China).

Techniques: Immunohistochemistry, Expressing, Transfection, Control, Western Blot, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Presenilin1 exerts antiproliferative effects by repressing the Wnt/β-catenin pathway in glioblastoma

doi: 10.1186/s12964-019-0501-9

Figure Lengend Snippet: a CCK-8 assays were performed in U87 cell when loss- or gain- function of Presenilin1, OD value were detected at day 1,2,3,5 and 7 and were showed as mean ± SD. b U251 cells down- or up-expressed of Presenilin1, then were used for CCK-8 assays to reveal the function of Presenilin1 in U251 cells. c Clone formation of the U87 cells after knockdown or over-expression of Presenilin1, histograms showing the clone formation in each group, Scale bars = 1 cm. d Representative image of clone formation in U251 cells after Sh-Presenilin1 or Lv-Presenilin1 treatment, Scale bars = 1 cm.* p < 0.05,** p < 0.01

Article Snippet: Rabbit anti-Presenilin1 monoclonal antibody was purchased from Proteintech (Wuhan, China).

Techniques: CCK-8 Assay, Knockdown, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Presenilin1 exerts antiproliferative effects by repressing the Wnt/β-catenin pathway in glioblastoma

doi: 10.1186/s12964-019-0501-9

Figure Lengend Snippet: a U87 and U251 cells were over-expressed of Presenilin1, then were used to flow cytometry assays to analyse the cell cycle. Histograms showing the percent of cell at G1, S and G2 phase. b Flow cytometry assays were applied to investigate the influence of down expression of Presenilin1 on cell cycle in U87 and U251 cells. c Edu staining to show the proliferation states of U87 and U251 cells after Presenilin1 were up-regulated, histograms showing the Edu positive cell ratio in each groups, Scale bars = 50um. d Representative Edu images and histograms to show the influence of Presenilin1 repression on glioma proliferation, Scale bars = 50um. e Western-blot assays to show the expression of C-myc, CDK6 and CyclinD1 when Presenilin1 down- or up-regulation in U87and U251 cell. * p < 0.05,** p < 0.01

Article Snippet: Rabbit anti-Presenilin1 monoclonal antibody was purchased from Proteintech (Wuhan, China).

Techniques: Flow Cytometry, Expressing, Staining, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Presenilin1 exerts antiproliferative effects by repressing the Wnt/β-catenin pathway in glioblastoma

doi: 10.1186/s12964-019-0501-9

Figure Lengend Snippet: a-c the correlation of Presenilin1 with β-catenin, CyclinD1 and C-myc from French-284-glioma dataset. d Western-blot assays to investigate the expression levels of p45-β-catenin, p33/37/41-β-catenin, total β-catenin, p9-GSK-3β and GSK-3β in U87 cells after down- or up-expression of Presenilin1. e-f The expression of β-catenin was detected by IF assays in U87 and U251 cells after down- or up-expression of Presenilin1. the co-localization mask and Pearson’s index were analyzed by Image J. scale bar = 20um, * p < 0.05,** p < 0.01

Article Snippet: Rabbit anti-Presenilin1 monoclonal antibody was purchased from Proteintech (Wuhan, China).

Techniques: Western Blot, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Presenilin1 exerts antiproliferative effects by repressing the Wnt/β-catenin pathway in glioblastoma

doi: 10.1186/s12964-019-0501-9

Figure Lengend Snippet: a-b Subcutaneous transplanting gliomas were utilized to analyse the function of Presenilin1 on glioma in vivo, tumor values were measured every-3 days and calculated to histogram. c Representative IHC images of Ki67 expression in 4 groups of subcutaneous glioma tissues, scale bar = 50um. d The expression of p45-β-catenin, p33/37/41-β-catenin and total β-catenin in 4 groups of subcutaneous glioma tissues were detected by IHC, scale bar = 50um. * p < 0.05,** p < 0.01. e Western-blot assays to investigate the expression levels of p45-β-catenin, p33/37/41-β-catenin, total β-catenin, p9-GSK-3β and GSK-3β in U87 cells after down- or up-expression of PSEN1

Article Snippet: Rabbit anti-Presenilin1 monoclonal antibody was purchased from Proteintech (Wuhan, China).

Techniques: In Vivo, Expressing, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Presenilin1 exerts antiproliferative effects by repressing the Wnt/β-catenin pathway in glioblastoma

doi: 10.1186/s12964-019-0501-9

Figure Lengend Snippet: a-d Intracranial xenografts of gliomas were established with GL2161 cells after up-regulation of Presenilin1, representative MRI image showing the the tumor size at 7 days and 19 days after transplating in each groups, scale bar = 5 mm. Histograms showing the tumor values at 7 days and 19 days after transplating in each groups. e The representative gross image of glioma after dissected from mice in each group. f IHC staining of Ki-67 to show the proliferation state of transplating glioma, Scale bar = 50um. g The survival curve of glioma-bearing mice was analyzed by Kaplan-Meier. h The schematic model: Presenilin1 exert anti-glioma function by increasing the phosphorylation of β-catenin and degradation, then arrest cell cycle at G1/S phase.* p < 0.05,** p < 0.01

Article Snippet: Rabbit anti-Presenilin1 monoclonal antibody was purchased from Proteintech (Wuhan, China).

Techniques: Immunohistochemistry, Phospho-proteomics