cfse dye  (Elabscience Biotechnology)

Journal: Journal of the Chinese Medical Association : JCMA

Article Title: Phenylalanyl-tRNA synthetase subunit beta downregulation by spi1 proto-oncogene modulates lung adenocarcinoma progression and immune microenvironment via mammalian target of rapamycin pathway

doi: 10.1097/JCMA.0000000000001286

Figure Lengend Snippet: FARSB promotes tumorigenesis and anti-tumor immunity in LUAD. A, qRT-PCR analysis of FARSB mRNA levels in different cell groups (si-NC, control group; si-FARSB, FARSB knockdown group). B, WB analysis of FARSB protein expression in various cell groups. C, CCK-8 assay evaluating the effect of FARSB knockdown on cell viability. D, EdU assay assessing the impact of FARSB knockdown on cell proliferation. E and F, Flow cytometry analysis of the effect of FARSB knockdown on cell apoptosis. G, WB analysis of PD-L1 (an immune checkpoint protein) expression levels. H, CFSE fluorescence staining assessing CD8 + T cell proliferation (CFSE dye dilution measures CD8 + T cell division). I-K, Flow cytometry analysis of IFN-γ (I), GZMB (J), and TNF-α (K) expressions in tumor-infiltrating CD8 + T cells; these cytokines reflect CD8 + T cell cytotoxic function. N = 3, *** p < 0.001 and **** p < 0.0001 denote statistical significance. CFSE = carboxyfluorescein succinimidyl ester; FARSB = phenylalanyl-tRNA synthetase subunit beta; GZMB = Granzyme B; IFN-γ = interferon-gamma; LUAD = lung adenocarcinoma; PD-L1 = programmed death-ligand 1; TNF-α = tumor necrosis factor-alpha; WB = western blot.

Article Snippet: Carboxyfluorescein succinimidyl ester (CFSE) dye (E-CK-A345, Elabscience, China) was diluted to the appropriate concentration and added to the CD8 + T cell suspension.

Techniques: Quantitative RT-PCR, Control, Knockdown, Expressing, CCK-8 Assay, EdU Assay, Flow Cytometry, Fluorescence, Staining, Western Blot

Journal: Journal of the Chinese Medical Association : JCMA

Article Title: Phenylalanyl-tRNA synthetase subunit beta downregulation by spi1 proto-oncogene modulates lung adenocarcinoma progression and immune microenvironment via mammalian target of rapamycin pathway

doi: 10.1097/JCMA.0000000000001286

Figure Lengend Snippet: FARSB Activates the mTOR signaling pathway, promoting LUAD tumorigenesis and anti-tumor immunity. A, GSEA analysis of enriched pathways associated with differential FARSB expression in LUAD. B, WB analysis of the expression of the pathway-related proteins p-mTOR, mTOR, p-p70S6K, p70S6K, p-4EBP1, and 4EBP1. C, CCK-8 assays evaluating cell viability. D, EdU assays assessing cell proliferation. E, Flow cytometry analysis of cell apoptosis. F, WB analysis of PD-L1 expression levels. G, CFSE fluorescence staining assessing CD8 + T cell proliferation. H-J, Flow cytometry analysis of IFN-γ (H), GZMB (I), and TNF-α (J) expressions in tumor-infiltrating CD8 + T cells; these cytokines reflect CD8 + T cell cytotoxic function. n = 3, ** p < 0.01, *** p < 0.001, **** p < 0.0001 denote statistical significance. CFSE = carboxyfluorescein succinimidyl ester; FARSB = phenylalanyl-tRNA synthetase subunit beta; GZMB = Granzyme B; IFN-γ = interferon-gamma; LUAD = lung adenocarcinoma; mTOR = mammalian target of rapamycin; ns = no significant differences; PD-L1 = programmed death-ligand 1; TNF-α = tumor necrosis factor alpha; WB = western blot.

Article Snippet: Carboxyfluorescein succinimidyl ester (CFSE) dye (E-CK-A345, Elabscience, China) was diluted to the appropriate concentration and added to the CD8 + T cell suspension.

Techniques: Expressing, CCK-8 Assay, Flow Cytometry, Fluorescence, Staining, Western Blot

Journal: Journal of the Chinese Medical Association : JCMA

Article Title: Phenylalanyl-tRNA synthetase subunit beta downregulation by spi1 proto-oncogene modulates lung adenocarcinoma progression and immune microenvironment via mammalian target of rapamycin pathway

doi: 10.1097/JCMA.0000000000001286

Figure Lengend Snippet: SPI1 Downregulates FARSB to modulate the mTOR signaling pathway, thus influencing LUAD progression and anti-tumor immunity. A, qRT-PCR analysis of FARSB and SPI1 mRNA levels in different cell groups. B, WB analysis of FARSB and SPI1 protein expression. C and D, WB analysis of mTOR signaling pathway-related proteins. E, CCK-8 assays evaluating cell viability. F, EdU assays assessing cell proliferation. G, Flow cytometry analysis of cell apoptosis. H, WB analysis of PD-L1 expression levels. I: CFSE fluorescence staining assessing CD8 + T cell proliferation. J-L: Flow cytometry analysis of IFN-γ (J), GZMB (K), and TNF-α (L) expressions in tumor-infiltrating CD8 + T cells; these cytokines reflect CD8+ T cell cytotoxic function. n = 3, *** p < 0.001, **** p < 0.0001 denote statistical significance. FARSB = phenylalanyl-tRNA synthetase subunit beta; GZMB = Granzyme B; IFN-γ = interferon-gamma; LUAD = lung adenocarcinoma; mTOR = Mammalian target of rapamycin; ns = no significant differences or significant correlation.; PD-L1 = programmed death-ligand 1; qRT-PCR = quantitative reverse transcription-polymerase chain reaction; TNF-α = tumor necrosis factor alpha; WB = western blot.

Article Snippet: Carboxyfluorescein succinimidyl ester (CFSE) dye (E-CK-A345, Elabscience, China) was diluted to the appropriate concentration and added to the CD8 + T cell suspension.

Techniques: Quantitative RT-PCR, Expressing, CCK-8 Assay, Flow Cytometry, Fluorescence, Staining, Reverse Transcription, Polymerase Chain Reaction, Western Blot