rabbit anti phosphorylated s6 ribosomal protein ps6  (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: iScience

Article Title: Fate plasticity of interneuron specification

doi: 10.1016/j.isci.2025.112295

Figure Lengend Snippet: Inhibition of the mTOR pathway in PNs reduces Pvalb specification (A) Experimental design: Chimeric cortical organoids were treated with either 250 nM Rapamycin or a vehicle (control) starting at the time of grafting and continuing for 14 days. (B) Representative images and quantification of phosphorylated ribosomal protein S6 (pS6) in both control and rapamycin-treated organoids at 14 DPG. White arrows indicate INs positive for both td-Tomato and pS6. Scale bar, 10 μm. n = 245 cells for control organoids and n = 342 cells for rapamycin-treated organoids. (C) Comparison of IN subtypes in control versus Rapamycin-treated organoids. Scale bar, 10 μm. n = 182 for control organoids and n = 210 for rapamycin-treated organoids. Unpaired parametric Student’s t test without Welch’s correction: ∗∗ = p < 0.01; ∗∗∗ = p < 0.001. Error bars represent SEM. See also Figures S13–S17 .

Article Snippet: Rabbit anti-S6 phosphorylated (pS6) (S235/236) , Cell Signaling , Cat#: 2211; RRID: AB_331679.

Techniques: Inhibition, Control, Comparison

Journal: iScience

Article Title: Fate plasticity of interneuron specification

doi: 10.1016/j.isci.2025.112295

Figure Lengend Snippet:

Article Snippet: Rabbit anti-S6 phosphorylated (pS6) (S235/236) , Cell Signaling , Cat#: 2211; RRID: AB_331679.

Techniques: Virus, Plasmid Preparation, Recombinant, Membrane, Imaging, RNA Sequencing, Software, Cell Culture, Modification, Knock-Out, Passaging

Journal: The FASEB Journal

Article Title: The roles of mTORC1 in parathyroid gland function in chronic kidney disease‐induced secondary hyperparathyroidism: Evidence from male genetic mouse models and clinical data

doi: 10.1096/fj.202401547RR

Figure Lengend Snippet: Phosphorylated ribosomal protein S6 levels are elevated in parathyroids of PT‐ Tsc1 −/− mice, but decreased in adenine‐rich high phosphorus‐induced CKD, unlike the increase in CKD control mice. (A) Immunofluorescence staining of parathyroid sections from control or adenine‐rich high‐phosphorus diet fed PT‐ Tsc1 +/+ and PT‐ Tsc1 −/− mice for nuclear SYTOX in green and phosphorylated ribosomal protein S6 (pS6) (Ser235/236) in red. Magnification ×10. (B) Quantification of staining intensity, measured using ImageJ software in this and one repeat experiment. Data are presented as mean ± SE . Statistical analysis was conducted using two‐way ANOVA and post‐hoc Tukey tests. * p < .05; ** p < .01; **** p < .0001.

Article Snippet: Immunostaining was conducted overnight at 4°C using a primary antibody targeting phosphorylated ribosomal protein S6 (pS6) (S235/236) at a dilution of 1:500 (Cell Signaling Technology, RRID:AB_2181037) and CaSR (1:1500, Novus Biologicals, Littleton, CO) diluted in Cas block (Zymed Laboratories, San Francisco, CA).

Techniques: Control, Immunofluorescence, Staining, Software

Journal: The FASEB Journal

Article Title: The roles of mTORC1 in parathyroid gland function in chronic kidney disease‐induced secondary hyperparathyroidism: Evidence from male genetic mouse models and clinical data

doi: 10.1096/fj.202401547rr

Figure Lengend Snippet: FIGURE 4 Phosphorylated ribosomal protein S6 levels are elevated in parathyroids of PT-Tsc1−/− mice, but decreased in adenine- rich high phosphorus-induced CKD, unlike the increase in CKD control mice. (A) Immunofluorescence staining of parathyroid sections from control or adenine-rich high-phosphorus diet fed PT-Tsc1+/+ and PT-Tsc1−/− mice for nuclear SYTOX in green and phosphorylated ribosomal protein S6 (pS6) (Ser235/236) in red. Magnification ×10. (B) Quantification of staining intensity, measured using ImageJ software in this and one repeat experiment. Data are presented as mean ± SE. Statistical analysis was conducted using two-way ANOVA and post-hoc Tukey tests. *p < .05; **p < .01; ****p < .0001.

Article Snippet: Immunostaining was conducted overnight at 4°C using a primary antibody targeting phosphorylated ribosomal protein S6 (pS6) (S235/236) at a dilution of 1:500 (Cell Signaling Technology, RRID:AB_2181037) and CaSR (1:1500, Novus Biologicals, Littleton, CO) diluted in Cas block (Zymed Laboratories, San Francisco, CA).

Techniques: Control, Immunofluorescence, Staining, Software

Journal: Cell proliferation

Article Title: The apoptotic and anti-proliferative effects of Neosetophomone B in T-cell acute lymphoblastic leukaemia via PI3K/AKT/mTOR pathway inhibition.

doi: 10.1111/cpr.13773

Figure Lengend Snippet: FIGURE 5 NSP-B's effect on Protein Kinase B/mammalian target of rapamycin (AKT/MTOR) signalling and apoptotic inhibitors in T cells. (A, B, E) Modulation of AKT/MTOR Pathway by NSP-B: Here, the impact of NSP-B on AKT (Ser473), MTOR (S2448), 4EBP1 and GSK3α/β phosphorylation and the expression of apoptotic inhibitors (XIAP, c-IAP1 and c-IAP2) is shown. Proteins from treated cells were resolved by SDS–PAGE and analysed by immunoblotting using specific antibodies. GAPDH and HSP60 were used as loading controls. (C) mTOR Activity Assessment: This panel depicts the intracellular levels of phosphorylated S6 ribosomal protein (pS6), a downstream target of mTOR, in cells treated with or without 2 μM NSB-P. Anti-pS6 monoclonal antibody was used for intracellular staining, with FACS histograms showing the mean fluorescence intensity (MFI). (D) Statistical analysis of pS6 MFI: A representative plot illustrates the pS6 MFI (mean ± SD) from two independent experiments, each conducted in triplicates. Statistical significance was calculated using an unpaired t-test (****p < 0.0001). NSP-B, Neosetophomone B.

Article Snippet: To assess mTOR activity, intracellular phosphorylated S6 ribosomal protein (pS6) levels were detected as described previously.28 Jurkat cells treated with NSP-B were fixed, permeabilized and stained with anti-pS6RP-Alexa Fluor 647 antibody (Ser235/236, clone 2F9, Cell Signaling Technologies).

Techniques: Phospho-proteomics, Expressing, SDS Page, Western Blot, Activity Assay, Staining, Fluorescence