anti ps6 s235  (Cell Signaling Technology Inc)

Journal: Cancer research communications

Article Title: Na + /H + -exchanger 1 enhances antitumor activity of engineered NK-92 natural killer cells

doi: 10.1158/2767-9764.crc-22-0270

Figure Lengend Snippet: (A) Liver weight of mice with doxycycline (DOX)-repressible c-Myc-driven HCC fed with regular or sodium bicarbonate (NaHCO3)-containing drinking water and treated with anti-PD1 antibody (iPD1) or isotype control (IgG). Each dot represents an individual mouse liver weight in the different groups, all determined upon euthanasia at twelve weeks after birth (N = 10, 43, 43, 30, and 31 animals for H2O + DOX, H2O + IgG, NaHCO3 + IgG, H2O + iPD1, and NaHCO3 + iPD1 groups, respectively; unpaired t-test). (B) Representative microscopic images of the mTORC1 target phospho-S6 ribosomal protein (pS6)-stained tumor slices. Scale: 100 μm (inset: 40 μm). (C) Quantification of pS6-positive objects in images in B (N = 5 whole-tumor scans from different animals, one-way ANOVA with multiple comparisons). * p < 0.05, *** p < 0.001.

Article Snippet: Cells were then stained with PE-Cy7-conjugated rabbit anti-pS6 (Ser235/Ser236) antibody (Cell Signaling Technology, 34411) or isotype control antibody (Cell Signaling Technology, 97492) at 1:50 dilution for 20 min on ice before analysis.

Techniques: Control, Staining

Journal: Cancer research communications

Article Title: Na + /H + -exchanger 1 enhances antitumor activity of engineered NK-92 natural killer cells

doi: 10.1158/2767-9764.crc-22-0270

Figure Lengend Snippet: (A) In vitro cytotoxicity of Torin1 (mTOR inhibitor)-treated NK-92 cells against the human melanoma cell line WM3629, relative to the untreated cells (E-T ratio = 3:1, N = 6 wells, one-way ANOVA with multiple comparisons versus untreated). (B) Representative immunoblots of the mTORC1 phosphorylation substrates phospho-p70 S6 kinase (pS6K) and phospho-S6 ribosomal protein (pS6) in constitutively active RHEB (RHEBN153T, hereafter referred to as RHEB)-overexpressing or empty vector (EV) control NK-92 cells treated with pH-controlled media for 6 or 24 hours. Endogenous RHEB is shown as the dim, lower band in the images, where the bright, upper band represents the overexpressed RHEB. (C) In vitro cytotoxicity of RHEB-expressing or EV control NK-92 cells against human melanoma cell lines WM3629 and WM4237 in pH-controlled media (E-T ratio = 3:1, N = 4 wells for each cell line, unpaired t-test). (D) Degranulation of RHEB-overexpressing or EV control NK-92 cells towards the human leukemia cell line K562 in pH-controlled media (E-T ratio = 1:2, N = 3 wells, unpaired t-test). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Cells were then stained with PE-Cy7-conjugated rabbit anti-pS6 (Ser235/Ser236) antibody (Cell Signaling Technology, 34411) or isotype control antibody (Cell Signaling Technology, 97492) at 1:50 dilution for 20 min on ice before analysis.

Techniques: In Vitro, Western Blot, Phospho-proteomics, Plasmid Preparation, Control, Expressing

Journal: Cancer research communications

Article Title: Na + /H + -exchanger 1 enhances antitumor activity of engineered NK-92 natural killer cells

doi: 10.1158/2767-9764.crc-22-0270

Figure Lengend Snippet: (A) Representative immunoblots assessing mTORC1 substrates pS6K and pS6 in NK-92 cells expressing constitutively active NHE1, NHE1-E262I, or EV treated with pH-controlled media for 6 or 24 hours. At the exposure shown, endogenous NHE1 is barely visible due to low expression. (B) Degranulation of trametinib (MEK inhibitor)-treated NK-92 cells against K562 cells, relative to untreated cells (E-T ratio = 1:2, N = 3 wells, one-way ANOVA with multiple comparisons versus untreated). (C) Representative immunoblots assessing phospho-p44/42 MAPK (pERK1/2) in NK-92 cells expressing NHE1, NHE1-E262I, or EV treated with pH-controlled media for 6 or 24 hours. At the exposure shown, endogenous NHE1 is barely visible due to low expression. (D) Percentage of NHE1-, NHE1-E262I-, or EV-expressing NK-92 cells positive for intracellular pERK1/2 overtime after interacting with K562 cells (E-T ratio = 1:1, N = 3 wells, two-way ANOVA with multiple comparisons). * p < 0.05, *** p < 0.001.

Article Snippet: Cells were then stained with PE-Cy7-conjugated rabbit anti-pS6 (Ser235/Ser236) antibody (Cell Signaling Technology, 34411) or isotype control antibody (Cell Signaling Technology, 97492) at 1:50 dilution for 20 min on ice before analysis.

Techniques: Western Blot, Expressing

Journal: Cell reports

Article Title: Methylation of dual-specificity phosphatase 4 controls cell differentiation

doi: 10.1016/j.celrep.2021.109421

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: PE Phospho-pS6 Ribosome Protein , Cell Signaling , Cat# 5316S; RRID:AB_10694989.

Techniques: Plasmid Preparation, Virus, Recombinant, Protease Inhibitor, Modification, Protein Purification, Western Blot, Membrane, Viability Assay, Polymer, cDNA Synthesis, Sequencing, Real-time Polymerase Chain Reaction, Software