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: Activating mTORC1 by RHEB enhances cytolytic activity of NK-92 cells. 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 pS6K and pS6 in constitutively active RHEB (RHEB N153T , hereafter referred to as RHEB)-overexpressing or 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 minutes on ice before analysis.
Techniques: Activity Assay, In Vitro, Western Blot, Phospho-proteomics, Control, Expressing