Journal: BMC Medical Genomics

Article Title: Pro-neural transcription factors as cancer markers

doi: 10.1186/1755-8794-1-17

Figure Lengend Snippet: NTR2 expression increases progressively during the cancer development whereas NT and Ascl1 expression is upregulated at the BPH-PIN transition . (a) Immunohistochemical analysis of prostate tissue microarrays comprising normal, BPH, PIN and adenocarcinomas. Sections were stained with anti-NT, anti-NTR2 or anti-Ascl1 antibodies. NTR2 immunoreactivity is most intense in PIN and carcinomas, in contrast NT and Ascl1 immunoreactivity is mostly observed in the carcinomas. (b-d) Histopathological score for each specific prostate tissue based on the immunoreactivity antibody staining.

Article Snippet: For NT (Sigma), NTR2 (Acris Antibodies GmbH) and Ascl1 (Aviva Systems Biology) antibodies, antigen retrieval was performed in Tris-EDTA at pH 9.0 in a microwave for 15 minutes.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Alcoholism, clinical and experimental research

Article Title: Heightened Exploratory Behavior Following Chronic Excessive Ethanol Drinking: Mediation by Neurotensin Receptor Type 2 in the Anterior Paraventricular Thalamus.

doi: 10.1111/acer.14406

Figure Lengend Snippet: Fig. 3. Chronic high ethanol (EtOH) drinkers exhibit reduced mRNA expression of neurotensin receptor type 2 (NTS2R) in the anterior paraventricular thalamus (aPVT), as assessed using quantitative real-time PCR (Experiment 3). (A) Comparison of the high EtOH drinkers (n = 6), low EtOH drinkers (n = 9), and water drinkers (controls) (n = 8) for mRNA levels of neurotensin (NTS), neurotensin receptor type 1 (NTS1R), and NTS2R in the aPVT fol- lowing 11 weeks of EtOH access (Week 12) during acute abstinence. Compared to low EtOH drinkers and water drinkers, the high EtOH drinkers showed significantly reduced expression of NTS2R mRNA in the aPVT. (B) Comparison of these same groups for the same measures in the posterior paraventricular thalamus revealed no group differences. Data are mean SEM, *p < 0.05.

Article Snippet: Tissue was immunotagged with polyclonal rabbit anti-NTS2R antibody (1:200, Cat # AP01326PU-N; Origene) and monoclonal mouse anti-NeuN antibody (1:1,000, Cat # ab104224; Abcam) and labeled with fluorescent goat anti-rabbit conjugated to AF488 (1:10,000, Cat # ab8245; Abcam) and goat anti-mouse conjugated to AF594 (1:500, Cat # A11005; Thermo Fisher Scientific).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Comparison

Journal: Alcoholism, clinical and experimental research

Article Title: Heightened Exploratory Behavior Following Chronic Excessive Ethanol Drinking: Mediation by Neurotensin Receptor Type 2 in the Anterior Paraventricular Thalamus.

doi: 10.1111/acer.14406

Figure Lengend Snippet: Fig. 4. Chronic high ethanol (EtOH) drinkers exhibit elevated protein levels of neurotensin receptor type 2 (NTS2R) in the anterior paraventricular tha- lamus (aPVT), as analyzed using Western blot (Experiment 4). (A) Comparison of the high EtOH drinkers (n = 6), low EtOH drinkers (n = 11), and water drinkers (controls) (n = 5) for protein levels of NTS2R in the aPVT following 11 weeks of EtOH access (Week 12) during acute abstinence. Compared to low EtOH drinkers and water drinkers, the high EtOH drinkers showed significantly elevated expression of NTS2R protein in the aPVT. (B) Comparison of these same groups for the same measures in the posterior paraventricular thalamus revealed no group differences. (C) Representative Western blot showing protein bands of NTS2R (green) and loading control GAPDH (red) in a water drinker (control), a low EtOH drinker, and a high EtOH drinker. (D) Original, full-size version of the Western blot, showing protein bands of NTS2R (green) and loading control GAPDH (red) in adult whole brain lysate for control (B), water drinkers (W), low EtOH drinkers (L), and high EtOH drinkers (H). Data are mean SEM, *p < 0.05.

Article Snippet: Tissue was immunotagged with polyclonal rabbit anti-NTS2R antibody (1:200, Cat # AP01326PU-N; Origene) and monoclonal mouse anti-NeuN antibody (1:1,000, Cat # ab104224; Abcam) and labeled with fluorescent goat anti-rabbit conjugated to AF488 (1:10,000, Cat # ab8245; Abcam) and goat anti-mouse conjugated to AF594 (1:500, Cat # A11005; Thermo Fisher Scientific).

Techniques: Western Blot, Comparison, Expressing, Control

Journal: Alcoholism, clinical and experimental research

Article Title: Heightened Exploratory Behavior Following Chronic Excessive Ethanol Drinking: Mediation by Neurotensin Receptor Type 2 in the Anterior Paraventricular Thalamus.

doi: 10.1111/acer.14406

Figure Lengend Snippet: Fig. 5. Elevated neurotensin receptor type 2 (NTS2R) signaling in the anterior paraventricular thalamus (aPVT) stimulates exploratory behavior (Experiment 5). (A) Average daily ethanol (EtOH) intake for low EtOH drinkers (n = 16) compared to high EtOH drinkers (n = 9) was significantly different at each week starting at Week 3. (B) Immunolabeling of NTS2R (green) and NeuN (neuronal nuclei; red), and nuclear staining by 40,6-diamidino-2- phenylindole (DAPI; blue) in the aPVT and posterior paraventricular thalamus (pPVT) shows expression of NTS2R in both neuronal cells (yellow) and nonneuronal cells (green). Images were taken at 10X magnification (left; scale bar: 100 µm) and 40X magnification (right; scale bar: 25 µm). (C) Sche- matic showing injection sites (black dots) for animals that received injections in the aPVT. Animals with a misplaced cannula, resulting in injections outside of the aPVT, were removed from analysis. Adapted from The Rat Brain, 5th edition, G. Paxinos and C. Watson, Copyright 2005, with permission from Elsevier. B = bregma. (D) Stimulating the NTS2R in the aPVT with the selective NTS2R agonist, JMV-431 (0.1 nmol or 1.0 nmol), compared to saline vehicle (0.3 µl), at Week 8 of EtOH access in low EtOH drinkers did not alter EtOH intake at any of the time periods measured during a 24-hour drinking session. (E) Stimulating the NTS2R in the aPVT with JMV-431 (1.0 nmol), compared to baseline or saline vehicle (0.3 µl), at Week 10 of EtOH access in low EtOH drinkers stimulated exploratory behavior in the light–dark box. Data are mean SEM, *p < 0.05.

Article Snippet: Tissue was immunotagged with polyclonal rabbit anti-NTS2R antibody (1:200, Cat # AP01326PU-N; Origene) and monoclonal mouse anti-NeuN antibody (1:1,000, Cat # ab104224; Abcam) and labeled with fluorescent goat anti-rabbit conjugated to AF488 (1:10,000, Cat # ab8245; Abcam) and goat anti-mouse conjugated to AF594 (1:500, Cat # A11005; Thermo Fisher Scientific).

Techniques: Immunolabeling, Staining, Expressing, Injection, Saline

Journal: Cell Discovery

Article Title: RhoA-mediated G 12 -G 13 signaling maintains muscle stem cell quiescence and prevents stem cell loss

doi: 10.1038/s41421-024-00696-7

Figure Lengend Snippet: a RT-qPCR analysis of Ednrb , Ntsr1 , and Ntsr2 expression in freshly isolated MuSCs (FSC) and activated stem cells (ASC) ( n = 3). b Representative images of immunostaining for ET B /NTS 1 /NTS 2 (red), PAX7 (green) and DAPI (blue) on FDB myofibers of WT mice at 0 h (upper panel) and 24 h (lower panel) of culturing. c The quantification of GPCR staining intensity ( n = 3). d , e Quantification of the ratios of PAX7 + KI67 + ( d ) and PAX7 + MYOD ‒ ( e ) MuSCs on FDB myofibers after 24 h culturing in the presence of DMSO, BQ788 or SR142948A ( n = 6). f , g Quantification of the ratio of PAX7 + KI67 + ( f ) and PAX7 + MYOD − ( g ) MuSCs on FDB myofibers after 24 h culturing in the presence of DMSO, ET-3, NT, ET-3 + BQ788, NT + SR142948A, or ET-3 + NT ( n = 6). h ‒ l Representative images ( h ) and quantification of PAX7 + ( i ), the ratio of PAX7 + KI67 + ( j ), and PAX7 + MYOD + ( k ) cells, and quantification of MuSCs outside the basal limina ( l ) on transverse sections of WT mice TA muscle after intramuscular injection of solvent, BQ788 (1 mg/kg bodyweight), or SR142948A (0.5 mg/kg bodyweight), respectively ( n = 3). The cartoon in h depicts the outline of the experimental design. The data represent means ± SEM, analyzed by unpaired t -test ( a , c ) and one-way ANOVA with Bonferroni’s multiple comparisons test ( d ‒ g and i ‒ l ). ND not detected. Scale bars: 5 µm in b , 10 µm in h .

Article Snippet: The whole length cDNA of human EDNRB and NTSR2 genes were cloned from EDNRB-Tango (# 66458, Addgene) and NTSR2-Tango (# 66458, Addgene) plasmids, respectively, and inserted into the pcDNA3.1(+) vector subsequently.

Techniques: Quantitative RT-PCR, Expressing, Isolation, Immunostaining, Staining, Injection, Solvent