ps6 rabbit polyclonal cell signaling 2215 ab 331682  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: The odor of a nontoxic tetrodotoxin analog, 5,6,11-trideoxytetrodotoxin, is detected by specific olfactory sensory neurons of the green spotted puffers but is processed in overlapping olfactory bulbar area with food odors

doi: 10.1101/2023.08.22.552781

Figure Lengend Snippet: Differential activation areas in the olfactory bulb by TDT and AAs. Immunohistochemistry employing anti-pS6 antibody on transverse OB sections exposed to the vehicle, AAs, and TDT. ( A ) Five vertical lines ( 1-5 ) in the sagittal brain diagram represent the rostral-caudal positions of transverse right OB sections, which are correspondingly exposed to the vehicle( B ), AAs( C ), and TDT( D ). Four fish examined for each condition showed similar results. Scale bar = 100 µm. The density quantification of pS6-positive neurons in the dorsomedial ( dm ) and ventrolateral ( vl ) part of each section was measured (n = 4; white squares in D5 correspond to an area of 10 4 µm 2 ). The rostral-caudal transition of pS6-immunopositive cell density labeled in the dm and vl part of the OB by odorant stimulations. Graphs are divided by the presented odors ( E ) or divided by dm or vl ( F ). The same lowercase letters denote non-significant differences and different letters represent a significant difference by two-factor repeated measure ANOVA with Holm correction (P < 0.05). Abbreviations : Tel , telencephalon; OB , olfactory bulb; d , dorsal; v , ventral; l , lateral; m , medial..

Article Snippet: Odorant-responsive cells were labeled with an anti-phosphorylated ribosomal protein S6 (pS6) rabbit polyclonal antibody (Ser244, Sers247 anti-pS6, 44-923G, Thermo Fisher Scientific, MA; 1:5,000), which was used in mouse OSNs ( ) and green spotted puffer OSNs , as a neural activity marker.

Techniques: Activation Assay, Immunohistochemistry, Labeling

Journal: iScience

Article Title: Trans-omic analysis reveals opposite metabolic dysregulation between feeding and fasting in liver associated with obesity

doi: 10.1016/j.isci.2024.109121

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-pS6 (Ser235/Ser236) , Cell Signaling Technology , #2211.

Techniques: Enzyme-linked Immunosorbent Assay, Glucose Assay, Sequencing, Software

Journal: BMC Gastroenterology

Article Title: Glucose metabolic upregulation via phosphorylation of S6 ribosomal protein affects tumor progression in distal cholangiocarcinoma

doi: 10.1186/s12876-023-02815-2

Figure Lengend Snippet: Representative images of immunohistochemistry for scoring of staining intensity for pS6 and GLUT1 in distal cholangiocarcinoma tissues

Article Snippet: We performed Immunohistochemistry with rabbit polyclonal antibodies against pS6 (S240/244) diluted 1:400 (Cell Signaling Technology, Danvers, MA, USA) and GLUT1 diluted 1:100 (Abcam, Tokyo, Japan).

Techniques: Immunohistochemistry, Staining

Journal: BMC Gastroenterology

Article Title: Glucose metabolic upregulation via phosphorylation of S6 ribosomal protein affects tumor progression in distal cholangiocarcinoma

doi: 10.1186/s12876-023-02815-2

Figure Lengend Snippet: Patient characteristics

Article Snippet: We performed Immunohistochemistry with rabbit polyclonal antibodies against pS6 (S240/244) diluted 1:400 (Cell Signaling Technology, Danvers, MA, USA) and GLUT1 diluted 1:100 (Abcam, Tokyo, Japan).

Techniques:

Journal: BMC Gastroenterology

Article Title: Glucose metabolic upregulation via phosphorylation of S6 ribosomal protein affects tumor progression in distal cholangiocarcinoma

doi: 10.1186/s12876-023-02815-2

Figure Lengend Snippet: Relationship between S6 phosphorylation or GLUT1 and patient characteristics

Article Snippet: We performed Immunohistochemistry with rabbit polyclonal antibodies against pS6 (S240/244) diluted 1:400 (Cell Signaling Technology, Danvers, MA, USA) and GLUT1 diluted 1:100 (Abcam, Tokyo, Japan).

Techniques: Phospho-proteomics

Journal: BMC Gastroenterology

Article Title: Glucose metabolic upregulation via phosphorylation of S6 ribosomal protein affects tumor progression in distal cholangiocarcinoma

doi: 10.1186/s12876-023-02815-2

Figure Lengend Snippet: a - c Scatter plots showing the relationship between GLUT1 and pS6, GLUT1 and SUV-max of FDG-PET, and pS6 and SUV-max of FDG-PET. d Representative images of immunohistochemistry using antibodies against GLUT1 and pS6 in distal cholangiocarcinoma tissues

Article Snippet: We performed Immunohistochemistry with rabbit polyclonal antibodies against pS6 (S240/244) diluted 1:400 (Cell Signaling Technology, Danvers, MA, USA) and GLUT1 diluted 1:100 (Abcam, Tokyo, Japan).

Techniques: Immunohistochemistry

Journal: Frontiers in Cell and Developmental Biology

Article Title: miR-514a promotes neuronal development in human iPSC-derived neurons

doi: 10.3389/fcell.2023.1096463

Figure Lengend Snippet: miR-514a upregulates mTOR signaling pathway in human iPSC-derived neurons. Neurons were infected with lentiviruses expressing miR-514a and cultured for 7 days. (A–C) Representative images of iPSC-derived neurons stained with anti-pS6 (red) antibody and Hoechst 33,258 (magenta) at 7 DIV. Scale bars, 100 µm. (D–F) pS6 positive neurons derived from three independent iPSC line, respectively, were quantified. mTOR activity was markedly increased in neurons expressed miR-514a. n = 3 independent experiments, at least 200 neurons were analyzed in each experiment. (G) Western blotting analysis showing the level of pS6 in cultured neurons at 7 DIV. (H) Quantification of pS6 level. n = 3 independent experiments. *** p < 0.001, ** p < 0.01, * p < 0.05. Abbreviations: pS6, phospho-S6 ribosomal protein.

Article Snippet: The following primary antibodies were used for western blotting: polyclonal rabbit antibody pS6 Ser 235/236 (1:500; Cell Signaling Technology) and monoclonal rabbit antibody β-actin (1:500; Cell Signaling Technology).

Techniques: Derivative Assay, Infection, Expressing, Cell Culture, Staining, Activity Assay, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: miR-514a promotes neuronal development in human iPSC-derived neurons

doi: 10.3389/fcell.2023.1096463

Figure Lengend Snippet: Blocking miR-514a using its sponges causes inhibition of neuronal development in iPSC-derived neurons. Neurons were infected with lentiviruses expressing miR-514a-5p sponge, miR-514a-3p sponge or scrambled shRNA as control and cultured for 7 days. (A) Diagrams of lentiviral vector constructs expressing sponges against miR-514a-5p and miR-514a-3p. The sponges were expressed under the U6 RNA polymerase Ⅲ promotor, and EGFP is expressed under the CMV promotor. (B) Representative images of iPSC-derived neurons (201B7) stained with anti-MAP2 (blue) antibody at 7 DIV. Scale bars, 50 µm. (C) Representative images of iPSC-derived neurons (201B7) stained with anti-MAP2 (blue) antibody at 7 DIV. Scale bars, 25 µm. (D) Representative images of iPSC-derived neurons (201B7) stained with anti-pS6 (red) antibody and Hoechst 33,258 (magenta) at 7 DIV. Scale bars, 100 µm. (E) Quantitative analysis of total dendrite length. n = 3 independent experiments, at least 100 neurons were analyzed in each experiment. (F) Quantitative analysis of neuronal soma size. n = 3 independent experiments, at least 100 neurons were analyzed in each experiment. (G) Quantitative analysis of pS6 positive cells. n = 3 independent experiments, at least 200 cells were analyzed in each experiment. n = 3 independent experiments. *** p < 0.001, ** p < 0.01, * p < 0.05. Abbreviations: CMV, cytomegalovirus; EGFP, enhanced green fluorescent protein; MAP2, microtubule-associated protein 2; pS6, phospho-S6 ribosomal protein; n. s., not significant.

Article Snippet: The following primary antibodies were used for western blotting: polyclonal rabbit antibody pS6 Ser 235/236 (1:500; Cell Signaling Technology) and monoclonal rabbit antibody β-actin (1:500; Cell Signaling Technology).

Techniques: Blocking Assay, Inhibition, Derivative Assay, Infection, Expressing, shRNA, Control, Cell Culture, Plasmid Preparation, Construct, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: miR-514a promotes neuronal development in human iPSC-derived neurons

doi: 10.3389/fcell.2023.1096463

Figure Lengend Snippet: Variation in miR-514a sequences among primate genomes affects the expression level of mature miR-514a. (A) Comparison of sequence of precursor-miR-514a across species. (B) Schematic representation of the mutation in pre-miR-514a-3. (the nucleotide position in the mutation is indicated with arrow). (C) Expression levels of mature miR-514a were measured using qRT-PCR in 201B7 iPSC-derived neurons infected with lentiviruses expressing native miR-514a, miR-514a mutant or scrambled shRNA as control. n = 3 independent experiments. (D) Representative images of iPSC-derived neurons stained with anti-MAP2 (blue) antibody at 7 DIV. Scale bars, 50 µm. (E) Representative images of iPSC-derived neurons stained with anti-MAP2 (blue) antibody at 7 DIV. Scale bars, 25 µm. (F) Representative images of iPSC-derived neurons stained with anti-pS6 (red) antibody and Hoechst 33,258 (magenta) at 7 DIV. Scale bars, 100 µm. (G) Quantitative analysis of total dendrite length. n = 3 independent experiments, at least 100 neurons were analyzed in each experiment. (H) Quantitative analysis of neuronal soma size. n = 3 independent experiments, at least 100 neurons were analyzed in each experiment. (I) Quantitative analysis of pS6 positive cells. n = 3 independent experiments, at least 200 cells were analyzed in each experiment. ** p < 0.01, * p < 0.05. Abbreviations: MAP2, microtubule-associated protein 2; pS6, phospho-S6 ribosomal protein; n. s., not significant.

Article Snippet: The following primary antibodies were used for western blotting: polyclonal rabbit antibody pS6 Ser 235/236 (1:500; Cell Signaling Technology) and monoclonal rabbit antibody β-actin (1:500; Cell Signaling Technology).

Techniques: Expressing, Comparison, Sequencing, Mutagenesis, Quantitative RT-PCR, Derivative Assay, Infection, shRNA, Control, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: miR-514a promotes neuronal development in human iPSC-derived neurons

doi: 10.3389/fcell.2023.1096463

Figure Lengend Snippet: miR-514a upregulates mTOR signaling pathway in human iPSC-derived neurons. Neurons were infected with lentiviruses expressing miR-514a and cultured for 7 days. (A–C) Representative images of iPSC-derived neurons stained with anti-pS6 (red) antibody and Hoechst 33,258 (magenta) at 7 DIV. Scale bars, 100 µm. (D–F) pS6 positive neurons derived from three independent iPSC line, respectively, were quantified. mTOR activity was markedly increased in neurons expressed miR-514a. n = 3 independent experiments, at least 200 neurons were analyzed in each experiment. (G) Western blotting analysis showing the level of pS6 in cultured neurons at 7 DIV. (H) Quantification of pS6 level. n = 3 independent experiments. *** p < 0.001, ** p < 0.01, * p < 0.05. Abbreviations: pS6, phospho-S6 ribosomal protein.

Article Snippet: The following primary antibodies were used for immunocytochemistry: monoclonal mouse antibody β-III tubulin (Tuj-1) (1:1,000; Sigma), polyclonal guinea pig antibody microtubule-associated protein 2 (MAP2) (1:1,000; Synaptic Systems, Goettingen, Germany) and polyclonal rabbit antibody phospho-S6 ribosomal protein (pS6) Ser 235/236 (1:1,000; Cell Signaling Technology, Danvers, MA, United States).

Techniques: Derivative Assay, Infection, Expressing, Cell Culture, Staining, Activity Assay, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: miR-514a promotes neuronal development in human iPSC-derived neurons

doi: 10.3389/fcell.2023.1096463

Figure Lengend Snippet: Blocking miR-514a using its sponges causes inhibition of neuronal development in iPSC-derived neurons. Neurons were infected with lentiviruses expressing miR-514a-5p sponge, miR-514a-3p sponge or scrambled shRNA as control and cultured for 7 days. (A) Diagrams of lentiviral vector constructs expressing sponges against miR-514a-5p and miR-514a-3p. The sponges were expressed under the U6 RNA polymerase Ⅲ promotor, and EGFP is expressed under the CMV promotor. (B) Representative images of iPSC-derived neurons (201B7) stained with anti-MAP2 (blue) antibody at 7 DIV. Scale bars, 50 µm. (C) Representative images of iPSC-derived neurons (201B7) stained with anti-MAP2 (blue) antibody at 7 DIV. Scale bars, 25 µm. (D) Representative images of iPSC-derived neurons (201B7) stained with anti-pS6 (red) antibody and Hoechst 33,258 (magenta) at 7 DIV. Scale bars, 100 µm. (E) Quantitative analysis of total dendrite length. n = 3 independent experiments, at least 100 neurons were analyzed in each experiment. (F) Quantitative analysis of neuronal soma size. n = 3 independent experiments, at least 100 neurons were analyzed in each experiment. (G) Quantitative analysis of pS6 positive cells. n = 3 independent experiments, at least 200 cells were analyzed in each experiment. n = 3 independent experiments. *** p < 0.001, ** p < 0.01, * p < 0.05. Abbreviations: CMV, cytomegalovirus; EGFP, enhanced green fluorescent protein; MAP2, microtubule-associated protein 2; pS6, phospho-S6 ribosomal protein; n. s., not significant.

Article Snippet: The following primary antibodies were used for immunocytochemistry: monoclonal mouse antibody β-III tubulin (Tuj-1) (1:1,000; Sigma), polyclonal guinea pig antibody microtubule-associated protein 2 (MAP2) (1:1,000; Synaptic Systems, Goettingen, Germany) and polyclonal rabbit antibody phospho-S6 ribosomal protein (pS6) Ser 235/236 (1:1,000; Cell Signaling Technology, Danvers, MA, United States).

Techniques: Blocking Assay, Inhibition, Derivative Assay, Infection, Expressing, shRNA, Control, Cell Culture, Plasmid Preparation, Construct, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: miR-514a promotes neuronal development in human iPSC-derived neurons

doi: 10.3389/fcell.2023.1096463

Figure Lengend Snippet: Variation in miR-514a sequences among primate genomes affects the expression level of mature miR-514a. (A) Comparison of sequence of precursor-miR-514a across species. (B) Schematic representation of the mutation in pre-miR-514a-3. (the nucleotide position in the mutation is indicated with arrow). (C) Expression levels of mature miR-514a were measured using qRT-PCR in 201B7 iPSC-derived neurons infected with lentiviruses expressing native miR-514a, miR-514a mutant or scrambled shRNA as control. n = 3 independent experiments. (D) Representative images of iPSC-derived neurons stained with anti-MAP2 (blue) antibody at 7 DIV. Scale bars, 50 µm. (E) Representative images of iPSC-derived neurons stained with anti-MAP2 (blue) antibody at 7 DIV. Scale bars, 25 µm. (F) Representative images of iPSC-derived neurons stained with anti-pS6 (red) antibody and Hoechst 33,258 (magenta) at 7 DIV. Scale bars, 100 µm. (G) Quantitative analysis of total dendrite length. n = 3 independent experiments, at least 100 neurons were analyzed in each experiment. (H) Quantitative analysis of neuronal soma size. n = 3 independent experiments, at least 100 neurons were analyzed in each experiment. (I) Quantitative analysis of pS6 positive cells. n = 3 independent experiments, at least 200 cells were analyzed in each experiment. ** p < 0.01, * p < 0.05. Abbreviations: MAP2, microtubule-associated protein 2; pS6, phospho-S6 ribosomal protein; n. s., not significant.

Article Snippet: The following primary antibodies were used for immunocytochemistry: monoclonal mouse antibody β-III tubulin (Tuj-1) (1:1,000; Sigma), polyclonal guinea pig antibody microtubule-associated protein 2 (MAP2) (1:1,000; Synaptic Systems, Goettingen, Germany) and polyclonal rabbit antibody phospho-S6 ribosomal protein (pS6) Ser 235/236 (1:1,000; Cell Signaling Technology, Danvers, MA, United States).

Techniques: Expressing, Comparison, Sequencing, Mutagenesis, Quantitative RT-PCR, Derivative Assay, Infection, shRNA, Control, Staining

Journal: Journal of cancer science and clinical therapeutics

Article Title: Systemic Delivery of a Dual PI3K/mTOR Inhibitor More Effective than Topical Delivery in Preventing Anal Carcinogenesis in an HPV Transgenic Mouse Model

doi: 10.26502/jcsct.5079153

Figure Lengend Snippet: Mean pAKT and pS6 protein expression, markers of PI3K and mTOR activity respectively, quantified via immunohistochemical staining sections of K14E6/E7 mice anal tissue. Mean ± standard deviation values are reported in the RawIntDen/Area (raw integrated density/area). The notation * represents statistical significance (p < 0.05). A, B. Mice that began treatment at 5 weeks of age/ normal anal histology. C, D. Mice that began treatment at 15 weeks of age/ low-grade anal dysplasia. E, F. Mice that began treatment at 25 weeks of age/ high-grade anal dysplasia.

Article Snippet: Sections were then stained with a monoclonal rabbit antibody against pAKT serine 473 (1:50 in 5% horse serum in PBS, antibody #3787; Cell Signaling Technology, Danvers, Massachusetts, USA) or a polyclonal rabbit antibody against pS6 ribosomal protein Ser235/236 (1:50 in 5% horse serum in PBS, antibody #2211; Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.

Techniques: Expressing, Activity Assay, Immunohistochemical staining, Staining, Standard Deviation