Journal: Cell Reports

Article Title: The Ubiquitin Ligase TRIP12 Limits PARP1 Trapping and Constrains PARP Inhibitor Efficiency

doi: 10.1016/j.celrep.2020.107985

Figure Lengend Snippet:

Article Snippet: GFP primary antibody (rabbit) , Torrey Pines Biolabs , Cat# TP401 071519; RRID: AB_10013661.

Techniques: Recombinant, Modification, Protease Inhibitor, Ligation, Labeling, Purification, Plasmid Preparation, Software

Journal: International Journal of Molecular Sciences

Article Title: Lgr5 Does Not Vary Throughout the Menstrual Cycle in Endometriotic Human Eutopic Endometrium

doi: 10.3390/ijms20010022

Figure Lengend Snippet: Immunofluorescence measurement of LGR5 expression throughout the menstrual cycle in eutopic endometrium. ( A ) Epithelial and stromal expression of LGR5 in control and endometriosis tissue in five phases of the menstrual cycle. ( B ) Total tissue expression across five phases of the menstrual cycle in control and endometriosis tissues. ( C ) Differences in LGR5 expression throughout the menstrual cycle between control and endometriosis tissue groups ( t -test for each phase; proliferative: p = 0.0242; secretory: p = 0.0424; menstruation: p = 0.0121). (Control: n = 24; endometriosis: n = 24). ( D ) Example of immunofluorescence of LGR5 in eutopic endometrium (early secretory phase). In blue: DAPI; in red: LGR5. * p < 0.05.

Article Snippet: Samples stained with monoclonal rabbit anti-LGR5 primary antibody (1 μL per million cells was used; BioNova Scientific, Fremont, CA, USA) and goat Alexa647 anti-rabbit secondary antibody (Invitrogen) at a 1:500 dilution.

Techniques: Immunofluorescence, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Lgr5 Does Not Vary Throughout the Menstrual Cycle in Endometriotic Human Eutopic Endometrium

doi: 10.3390/ijms20010022

Figure Lengend Snippet: In vitro analysis of LGR5 expression throughout the menstrual cycle in endometrial stromal fibroblast primary culture after treatment (E 2 and E 2 P 4 for six and six more days, respectively). ( A ) Left panels show expression of CYR61 (marker of the proliferative phase) and DKK1 (marker of the secretory phase) in the control group ( n = 4); right panels show expression of CYR61 and DKK1 in the endometriosis group ( n = 3). ( B ) Upper panel shows expression of LGR5 in proliferative (E 2 ) and secretory (E 2 P 4 ) phases of the control group; lower panel shows LGR5 expression in both phases of the endometriosis group (E 2 : estradiol; P 4 : progesterone).

Article Snippet: Samples stained with monoclonal rabbit anti-LGR5 primary antibody (1 μL per million cells was used; BioNova Scientific, Fremont, CA, USA) and goat Alexa647 anti-rabbit secondary antibody (Invitrogen) at a 1:500 dilution.

Techniques: In Vitro, Expressing, Marker, Control

Journal: International Journal of Molecular Sciences

Article Title: Lgr5 Does Not Vary Throughout the Menstrual Cycle in Endometriotic Human Eutopic Endometrium

doi: 10.3390/ijms20010022

Figure Lengend Snippet: Percentage of FACS-sorted LGR5 + cells. ( A ) Percentages of LGR5 + cells in different phases of the cycle (proliferative: n = 9; secretory: n = 4; ovulatory: n = 12; ND: n = 9). ( B ) Percentage of LGR5 + cells in eutopic endometrium of different types of endometriosis and controls (ovarian: n = 8; pelvic: n = 3; adenomyosis: n = 4; DIE: n = 10; control: n = 12). ( C ) Percentage of LGR5 + cells identified in eutopic endometrium of women with and without endometriosis (control: n = 12; endometriosis: n = 25). ( D ) Differences in LGR5 + cells between eutopic and ectopic endometrium. Left panel: ovarian endometriosis (eutopic: n = 8; ectopic: n = 4; p = 0.0286). Right panel: DIE (eutopic: n = 10; ectopic: n = 6; p = 0.0411). ( E ) Difference between ovarian and DIE ectopic endometrium (ovarian: n = 4; DIE: n = 6; p = 0.0381). ND: non-determined; DIE: deep infiltrating endometriosis. * p < 0.05, ** p < 0.03.

Article Snippet: Samples stained with monoclonal rabbit anti-LGR5 primary antibody (1 μL per million cells was used; BioNova Scientific, Fremont, CA, USA) and goat Alexa647 anti-rabbit secondary antibody (Invitrogen) at a 1:500 dilution.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Lgr5 Does Not Vary Throughout the Menstrual Cycle in Endometriotic Human Eutopic Endometrium

doi: 10.3390/ijms20010022

Figure Lengend Snippet: Primers used for RT-qPCR.

Article Snippet: Samples stained with monoclonal rabbit anti-LGR5 primary antibody (1 μL per million cells was used; BioNova Scientific, Fremont, CA, USA) and goat Alexa647 anti-rabbit secondary antibody (Invitrogen) at a 1:500 dilution.

Techniques: Sequencing

Journal: Journal of Heredity

Article Title: Differences in Cell Proliferation and Craniofacial Phenotype of Closely Related Species in the Pupfish Genus Cyprinodon

doi: 10.1093/jhered/esz074

Figure Lengend Snippet: Immunofluorescence light sheet imaging visualizes number and location of mitotic cells as indicated by assay for pH3 in the heads of hatching-age Cyprinodon. (A) Regions of the head analyzed for proliferating cells. (B) 3D reconstructions in lateral view of the head for the DAPI channel (gray, stains nuclei), the pH3 channel (red, dividing cells), and both channels merged. Note that pH3 positive cells (red dots) are mainly localized to ventral structures as shown in lateral view. (C, D) Single 2D images in (C) frontal view and (D) lateral view show pH3 cells (red dots) localizing to epithelial and mesenchyme tissues surrounding cartilage elements as exemplified by the ceratohyal. Cartilage cells are identifiable in the DAPI channel (gray) by their widely spaced nuclei indicative of large cuboidal cells. Cartilage elements can be identified by shape, and are outlined by the brightly labeled perichondrium, a dense layer of mesenchyme cells that surrounds cartilage elements. Inset shows region outlined by white box and arrows point to pH3 positive cells in perichondrium. Note also the clusters of pH3 positive cells in jaws (arrow head), especially around lateral edge of jaws as seen in panel C. Labels: bh, basihyal cartilage; br, brain; ch, ceratohyal cartilage; ey, eye; ga, gill arches; lj, lower jaw; pa, pharynx; pf, pectoral fin; uj, upper jaw.

Article Snippet: Cells were labeled for pH3 with primary rabbit anti-pH3 polyclonal antibody (MyBioSource, MBS2517807), and secondary antibody Alexa Fluor 568 goat anti-Rabbit (Thermo Fisher, A11011).

Techniques: Immunofluorescence, Imaging, Labeling

Journal: Journal of Heredity

Article Title: Differences in Cell Proliferation and Craniofacial Phenotype of Closely Related Species in the Pupfish Genus Cyprinodon

doi: 10.1093/jhered/esz074

Figure Lengend Snippet: Cell proliferation varies among species of Cyprinodon in different regions of the head at hatching. Shown are number of pH3 positive cells relative to either surface area of sampled tissue (A) or volume of sampled tissue (B) for the 3 regions of the head sampled plus the index of relative jaw proliferation (see text). Plotted are values for each sample and boxplots. Samples sizes are snail-eater N = 30, omnivore N = 28, scale-biter N = 19. Significance for all post hoc pairwise comparisons (Tukey) is shown above boxplots and corresponds to Supplementary Table S3. Note that levels of proliferation significantly vary between species when sampling the entire head (e.g., head region standardized to surface area) or regions of the head posterior to the jaws (post-jaw subset), but proliferation does not vary in the jaws (jaw subset). ns, not significant. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Cells were labeled for pH3 with primary rabbit anti-pH3 polyclonal antibody (MyBioSource, MBS2517807), and secondary antibody Alexa Fluor 568 goat anti-Rabbit (Thermo Fisher, A11011).

Techniques: Sampling

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