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PTRF/Cavin-1 as a Novel RNABinding Protein Expedites the NF-kB/PD-L1 Axis by Stabilizing lncRNA NEAT1, Contributing to Tumorigenesis and Immune Evasion in Glioblastoma
285 人阅读发布时间：2022-05-18 10:33
T Cell Cytotoxicity Assay
Control and PTRF overexpressing TBD0220 cells were separately
transfected with siRNA for NEAT1 in DMEM/F12 medium.
Human peripheral blood mononuclear cells (PBMC; CBP-002,
StemEry) were maintained in DMEM/F12 medium. The PBMC
cells were activated with 100 ng/mL CD3 antibody (317325,
BioLegend), 100 ng/mL CD28 antibody (302933, BioLegend),
and 10 ng/mL IL2 (589102, BioLegend) for 24 h. Then, activated
PBMC cells were incubated with TBD0220 cells at 10:1 ratio in
the presence of caspase-3/7 green detection reagent (C10723,
Thermo Fisher, 1:1000) at 37°C for 12 hours according to the
manufacturer’s protocol. The cell nuclei were visualized by
staining with DAPI. All the images were observed using FV-
1200 laser scanning confocal microscope (Olympus
Corporation, Japan)
Control and PTRF overexpressing TBD0220 cells were separately
transfected with siRNA for NEAT1 in DMEM/F12 medium.
Human peripheral blood mononuclear cells (PBMC; CBP-002,
StemEry) were maintained in DMEM/F12 medium. The PBMC
cells were activated with 100 ng/mL CD3 antibody (317325,
BioLegend), 100 ng/mL CD28 antibody (302933, BioLegend),
and 10 ng/mL IL2 (589102, BioLegend) for 24 h. Then, activated
PBMC cells were incubated with TBD0220 cells at 10:1 ratio in
the presence of caspase-3/7 green detection reagent (C10723,
Thermo Fisher, 1:1000) at 37°C for 12 hours according to the
manufacturer’s protocol. The cell nuclei were visualized by
staining with DAPI. All the images were observed using FV-
1200 laser scanning confocal microscope (Olympus
Corporation, Japan)