ps6 rb  (Cell Signaling Technology Inc)

Journal: NPJ Parkinson's Disease

Article Title: Impulsive-compulsive behaviours and striatal neuroactivity in mildly parkinsonian rats under D2/3 agonist and L-DOPA treatment

doi: 10.1038/s41531-025-00996-z

Figure Lengend Snippet: a pS6-immunopositive (pS6 + ) cell counts in the whole striatum under different treatments ( n = 69). Two-factor ANOVA was followed by Tukey’s post hoc test for the overall treatment effect. F(treatment) 3,61 = 8.9, p < 0.001; F(lesion) 1,61 = 0.9, p = 0.339; F(interaction) 3,61 = 1.7, p = 0.183. b pS6 + cell counts in the whole striatum and the denervated area in 6-OHDA-lesioned animals under different treatments ( n = 35). Two-factor repeated measurement ANOVA was followed by Tukey’s post hoc test for pairwise comparisons within one treatment or lesion type. F(treatment) 3,31 = 3.3, p = 0.033; F(area) 1,31 = 210.0, p < 0.001; F(animal) 31,31 = 15.9, p < 0.001; F(area x treatment) 3,31 = 8.4, p < 0.001. c Representative immunohistochemical stainings of pS6 + cells in the striatum developed using 3,3’ - diaminobenzidine (DAB). Shown are high magnification images of the intact and the lesioned area of one example animal per treatment. Scale bar: 30 μm. d Representative immunohistochemical stainings of pS6 + cell distributions in the striatum developed using DAB from one example animal per experimental group. Scale bar: 300 μm. Symbols of statistical significance: a = p < 0.05 vs Saline; b = p < 0.05 vs LD24; c = p < 0.05 vs R2.5; bracket = p < 0.05 for Total vs Lesioned area within the same treatment.

Article Snippet: For quantifying the extent of striatal dopaminergic denervation and counting of pS6 + cells, bright-field immunohistochemistry was performed using a primary antibody against TH (rabbit anti-TH, Pel-Freez P40101 , 1:1000) and phosphorylated pS6, respectively (monoclonal rabbit anti Ser235/236-phospho-S6, Cell Signaling #2211, 1:200).

Techniques: Immunohistochemical staining, Saline

Journal: NPJ Parkinson's Disease

Article Title: Impulsive-compulsive behaviours and striatal neuroactivity in mildly parkinsonian rats under D2/3 agonist and L-DOPA treatment

doi: 10.1038/s41531-025-00996-z

Figure Lengend Snippet: a –c Main principal components (PCs) identified in a principal component analysis of 2D histograms of pS6 + cell distributions in the striatum. Colour scale shows local variance, V max = maximal variance. Covariance is present in pixels with the same variance sign (positive: red, or negative: blue) and antivariance is present in pixels with opposite variance signs (red vs blue). a PC1. b PC2. c Inverted PC3. a’ –c’ Coefficients of main PCs as indices of the expression level of each covariance pattern for pS6 + cell distributions in different experimental groups. Two-factor ANOVAs were followed by Tukey’s post hoc test for pairwise comparisons within one treatment or lesion type; n(independent animals)=69, n(sections)=411. a’ Coefficient of PC1. F(treatment) 3,61 = 28.3, p < 0.001; F(lesion) 1,61 = 5.3, p = 0.024; F(interaction) 3,61 = 6.1, p = 0.001. b’ Coefficient of PC2. F(treatment) 3,61 = 1.6, p = 0.191; F(lesion) 1,61 = 62.3, p < 0.001; F(interaction) 3,61 = 21.7, p < 0.001. c’ Coefficient of inverted PC3. F(treatment) 3,61 = 10.0, p < 0.001; F(lesion) 1,61 = 2.1, p = 0.155; F(interaction) 3,61 = 7.3, p < 0.001. Symbols of statistical significance: a = p < 0.05 vs Saline within the same lesion type; b = p < 0.05 vs LD24 within the same lesion type; c = p < 0.05 vs R2.5 within the same lesion type; bracket = p < 0.05 for Sham vs 6-OHDA within the same treatment.

Article Snippet: For quantifying the extent of striatal dopaminergic denervation and counting of pS6 + cells, bright-field immunohistochemistry was performed using a primary antibody against TH (rabbit anti-TH, Pel-Freez P40101 , 1:1000) and phosphorylated pS6, respectively (monoclonal rabbit anti Ser235/236-phospho-S6, Cell Signaling #2211, 1:200).

Techniques: Expressing, Saline

Journal: Proceedings. Biological sciences

Article Title: Behavioural and neural correlates of social hierarchy formation in a sex-changing fish.

doi: 10.1098/rspb.2024.2097

Figure Lengend Snippet: Figure 3. Neural activation patterns after social disruption. Representative images for each brain area are depicted on the left (i; rank 2) and middle (ii; ranks 3−5) in each panel. Boxplots of pS6 counts are depicted on the right of each panel, with 2c and 3−5c representing the controls (iii). (A) The medial part of the dorsal telencephalon (Dm), with magnification of the insets to the right of each panel (scale bar of insets = 20 µm). (B) The dorsal part of the ventral telencephalon (Vd). (C) The ventral part of the ventral telencephalon (Vv). (D) The supracommissural nucleus of the ventral telencephalon (Vs). (E) The central nucleus of the telencephalic area (Vc). (F) the preoptic area (POA). (G) the ventral tuberal region of the hypothalamus (vTn). (H) the periaqueductal gray (PAG). (I) the periventricular nucleus of the posterior tuberculum (TPp). (J) The anterior tuberal nucleus (aTn; figure 4J). Scale bar = 100 µm. (K) Overview of brain regions with significantly different pS6 counts in rank 2 fish compared to other ranks.

Article Snippet: Non-specific binding was blocked (1% bovine serum albumin, 0.3% Triton-X and 5.0% normal goat serum made in 1 × PBS) for 2 h at room temperature, and slides were incubated with primary pS6 antibody (1 : 1500; Cell Signaling Technologies pS6 ribosomal protein S235/236 rabbit monoclonal antibody no. 4858) overnight at 4°C.

Techniques: Activation Assay, Disruption

Journal: Proceedings. Biological sciences

Article Title: Behavioural and neural correlates of social hierarchy formation in a sex-changing fish.

doi: 10.1098/rspb.2024.2097

Figure Lengend Snippet: Figure 3. Neural activation patterns after social disruption. Representative images for each brain area are depicted on the left (i; rank 2) and middle (ii; ranks 3−5) in each panel. Boxplots of pS6 counts are depicted on the right of each panel, with 2c and 3−5c representing the controls (iii). (A) The medial part of the dorsal telencephalon (Dm), with magnification of the insets to the right of each panel (scale bar of insets = 20 µm). (B) The dorsal part of the ventral telencephalon (Vd). (C) The ventral part of the ventral telencephalon (Vv). (D) The supracommissural nucleus of the ventral telencephalon (Vs). (E) The central nucleus of the telencephalic area (Vc). (F) the preoptic area (POA). (G) the ventral tuberal region of the hypothalamus (vTn). (H) the periaqueductal gray (PAG). (I) the periventricular nucleus of the posterior tuberculum (TPp). (J) The anterior tuberal nucleus (aTn; figure 4J). Scale bar = 100 µm. (K) Overview of brain regions with significantly different pS6 counts in rank 2 fish compared to other ranks.

Article Snippet: Specificity of the staining was confirmed through omission of either the primary antibody (electronic supplementary material, figure S3B) or the secondary antibody (electronic supplementary material, figure S3C) and through the use of a pS6 blocking peptide (Cell Signalling no. 1220; electronic supplementary material, figure S3D).

Techniques: Activation Assay, Disruption

Journal: bioRxiv

Article Title: The Functional and Genetic Architecture of Olfaction in Deer Mice

doi: 10.1101/2025.05.12.653383

Figure Lengend Snippet: ( A ) Mapping of cholesterol-derivatives detection on Vmn1r gene tree. Clades including specific receptors detecting each steroid class are indicated by dots. ( B ) Vmn1r F and ( C ) Vmn1r J receptor gene trees including homologs from mouse (crimson), rat (steel blue) and deer mouse (golden apricot). ( D ) Heatmap showing the percentage of protein identity for mouse Vmn1r237, Vmn1r89 and Vmn1r85 and their deer mouse nearest neighbor in the phylogeny. ( E ) Example of AlphaFold2 (AF2) predicted structure for a Vmn1r. The predicted confidence metrics (plDDT) indicate an overall high local accuracy in the models. ( F ) Comparison of AF2 predicted structures. For each pair, the Root Mean Square Deviation (RMSD) value is calculated based on pruned atom pairs in ChimeraX. ( G ) Summary of the detection of selected steroids by Vmn1r237, Vmn1r89 and Vmn1r85 in mouse. ( H ) Immunofluorescence with anti-pS6 (240/244) antibody (green) and in situ hybridization with RNA probe (red) after deer mouse VNO stimulation by individual steroids. Arrowheads mark colocalization between pS6 and receptor signals. The estriol E2734 only activates Vmn1rf7 whereas the androgen A7864 activates neither Vmn1rf7 nor Vmn1rj1/2. Vmn1rf7 is activated by most tested estradiols (E1100, E0893, E0588) but not by E1050. The estradiols E1050 and E1100 activate Vmn1rj1/2-expressing neurons. Overall, Vmn1rf7 in deer mouse shows a pattern highly similar to its mouse homolog Vmn1r237 (V1Rf3). Likewise, deer mouse Vmn1rj1/2 and mouse Vmn1r85 exhibit identical profiles.

Article Snippet: 500 μL of blocking buffer containing 1× PBS / 5 % normal goat serum /0.1 % Tween 20 was applied on the slides for 1 h at room temperature. pS6 antibody solution (anti-phosphorylated ribosomal protein S6 rabbit polyclonal (Ser240/244) Antibody, Cell Signaling #2215, at 1/250 dilution in antibody dilution buffer containing 1×PBS/ 1%BSA/0.1 % Tween 20) was applied on the slides, which were covered with parafilm and incubated in a sealed chamber for 40 h at 4°C.

Techniques: Comparison, Immunofluorescence, In Situ Hybridization, Expressing

Journal: bioRxiv

Article Title: The Functional and Genetic Architecture of Olfaction in Deer Mice

doi: 10.1101/2025.05.12.653383

Figure Lengend Snippet: ( A ) Coronal section of the vomeronasal organ from a deer mouse harvested post-exposure to snake feces for 1 hour. Activated olfactory sensory neurons in the sensory epithelium are identified using an antibody against pS6 (240/244) and stained with a fluorescently-labeled secondary antibody (pseudo-colored in green). Green puncta in the sensory epithelium correspond to neurons responding to cues present in the stimulus. ( B ) Tissue section harvested from a deer mouse exposed to clean bedding for 1 hour as control. Low background level of pS6 activity in the sensory epithelium underlines the suitability of pS6 (240/244) as an activity-dependent marker.

Article Snippet: 500 μL of blocking buffer containing 1× PBS / 5 % normal goat serum /0.1 % Tween 20 was applied on the slides for 1 h at room temperature. pS6 antibody solution (anti-phosphorylated ribosomal protein S6 rabbit polyclonal (Ser240/244) Antibody, Cell Signaling #2215, at 1/250 dilution in antibody dilution buffer containing 1×PBS/ 1%BSA/0.1 % Tween 20) was applied on the slides, which were covered with parafilm and incubated in a sealed chamber for 40 h at 4°C.

Techniques: Staining, Labeling, Control, Activity Assay, Marker