flow cytometry methods  (CellCarta)

Journal: Bioactive Materials

Article Title: STAT3-specific nanocarrier for shRNA/drug dual delivery and tumor synergistic therapy

doi: 10.1016/j.bioactmat.2024.07.010

Figure Lengend Snippet: (A) IC 50 results of DOX·HCl、DOX/VCPH、DOX/VCPH/pDNA nanoparticles with different DOX concentrations against A549 cells. (n = 6) (B) The relative inhibition rate of A549 cells transfected with different nanoparticles (* P < 0.05, n = 6). (C) The Calcein/PI staining images of A549 cells with different nanoparticles. (n = 3) (D) The flow cytometry result of A549 cells with different nanoparticles. (n = 3) (E) GFP-positive rate detected by flow cytometry of A549 cells transfected with different nanocomposite particles(compared with the negative control group, ** P < 0.01, compared with the positive control group, ## P < 0.01, compared between DOX/VCPH/pDNA and VCPH/pDNA, △△ P < 0.01, n = 6). (F) Laser confocal scanning microscopy of A549 cells transfected with different nanocomposite particles. (n = 3).

Article Snippet: After 4 h of cultivation, the complexes were removed and replaced with fresh DMEM, after which the well plates were incubated at 37 °C for another 48 h. The results were obtained using a fluorescence microscope (Leica, Germany) and by using the flow cytometry method (FACE Vantage, BD, USA).

Techniques: Inhibition, Transfection, Staining, Flow Cytometry, Negative Control, Positive Control, Confocal Laser Scanning Microscopy

Journal: Cancer Discovery

Article Title: CD8-Targeted IL2 Unleashes Tumor-Specific Immunity in Human Cancer Tissue by Reviving the Dysfunctional T-cell Pool

doi: 10.1158/2159-8290.cd-23-1263

Figure Lengend Snippet: Figure 1. CD8–IL2 selectively and potently activates CD8+ T cells in human tumor fragments. A, Overview of the PDTF platform and analysis strategy (created with BioRender.com). B, Representative flow cytometry plots displaying markers of proliferation (Ki-67), cytotoxicity (granzyme B), and activation (PD-1, CD137) in intratumoral CD8+ T cells from PDTFs that were left untreated or treated with ex vivo CD8–IL2 (RE098). C, Quantification of activation markers on total intratumoral CD8+ T cells in untreated or CD8–IL2-treated PDTFs measured by flow cytometry (n = 23). (continued on next page)

Article Snippet: Afterward, the samples were subjected to the flow cytometry staining procedure as described above and measured on an Aurora (Cytek Bio) analyzer.

Techniques: Flow Cytometry, Activation Assay, Ex Vivo

Journal: Cancer Discovery

Article Title: CD8-Targeted IL2 Unleashes Tumor-Specific Immunity in Human Cancer Tissue by Reviving the Dysfunctional T-cell Pool

doi: 10.1158/2159-8290.cd-23-1263

Figure Lengend Snippet: Figure 1. (Continued) D, Quantification of intracellular IFNγ in total intratumoral CD8+ T cells in untreated or CD8–IL2-treated tumor digests meas ured by flow cytometry (n = 5). E, Representative gating for PD-1+CD39+ (late dysfunctional) and PD-1+CD39− (early dysfunctional cells) CD8+ T cells. F, Same analysis as in C but separated for PD-1+CD39+ (late dysfunctional) and PD-1+CD39− (early dysfunctional cells). ****, P < 0.0001; ***, P < 0.001; **, P < 0.01 by two-tailed Wilcoxon test (C, D). ***, P < 0.001; **, P < 0.01 by the Friedman test corrected for multiple comparisons (F). Only significant comparisons are shown.

Article Snippet: Afterward, the samples were subjected to the flow cytometry staining procedure as described above and measured on an Aurora (Cytek Bio) analyzer.

Techniques: Flow Cytometry, Two Tailed Test

Journal: Cancer Discovery

Article Title: CD8-Targeted IL2 Unleashes Tumor-Specific Immunity in Human Cancer Tissue by Reviving the Dysfunctional T-cell Pool

doi: 10.1158/2159-8290.cd-23-1263

Figure Lengend Snippet: Figure 2. CD8–IL2 induces immunologic responses in a subset of tumors. A, Heat map displaying normalized delta values (CD8–IL2 condition − untreated condition) of 24 soluble mediators secreted by PDTFs, ordered according to unsupervised hierarchical clustering (n = 23). B, Examples of T-cell effector cytokines (IFNγ, TNFα), cytotoxic mediators (granzyme B), and chemokines (CXCL9, CXCL10, and CXCL11) in unstimulated and CD8–IL2 stimulated PDTFs in ex vivo responding (R) and nonresponding (NR) tumors. C, Separation of tumors by cumulative z-scores of soluble mediators in CD8–IL2 R and NR tumors. D, Baseline infiltration of dysfunctional (PD-1+CD39+CD103+) and memory (PD-1+IL7R+) CD8+ T cells and CXCL13 expression in CD8–IL2-R and NR tumors measured by flow cytometry. E, CD8+ T-cell activation markers measured by flow cytometry separately plotted for R and NR tumors. ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05 by two-tailed Wilcoxon test (B–E). Only significant comparisons are shown.

Article Snippet: Afterward, the samples were subjected to the flow cytometry staining procedure as described above and measured on an Aurora (Cytek Bio) analyzer.

Techniques: Ex Vivo, Expressing, Flow Cytometry, Activation Assay, Two Tailed Test

Journal: Cancer Discovery

Article Title: CD8-Targeted IL2 Unleashes Tumor-Specific Immunity in Human Cancer Tissue by Reviving the Dysfunctional T-cell Pool

doi: 10.1158/2159-8290.cd-23-1263

Figure Lengend Snippet: Figure 3. TCR signaling is required for a functional immune response to CD8–IL2. A, Schematic overview of Lck inhibition (LCKi) in the context of CD8–IL2 treatment (created with BioRender.com). B, Normalized delta values (CD8–IL2 or CD8–IL2 + LCKi condition − untreated condition) of soluble mediators secreted by PDTFs. Tumor samples were selected based on prior response to CD8–IL2 (as shown in Fig. 2A) and material availability (n = 8). C, Cumulative z-scores of soluble mediators in CD8–IL2-responding tumors when treated with either CD8–IL2 or CD8–IL2 + LCKi. D, Correlation of log2 fold changes (LOG2FC) of soluble mediators induced by either CD8–IL2 and CD8–IL2 + LCKi versus the untreated condition. E, Intratumoral CD8+ T-cell activation of CD8–IL2-responding tumors upon CD8–IL2 and CD8–IL2 + LCKi treatment measured by flow cytometry. F, Intracellular expression of ranzyme B and (G) secreted soluble Granzyme B and IFNγ in human PBMCs upon 5-day incubation with decreasing concentrations of anti-CD3 in the absence and presence of CD8–IL2 (n = 4). **, P < 0.01 by two-tailed Mann–Whitney U test (C). ****, P < 0.0001; ***, P < 0.001; **, P < 0.01 by the Friedman test corrected for multiple comparisons (E).

Article Snippet: Afterward, the samples were subjected to the flow cytometry staining procedure as described above and measured on an Aurora (Cytek Bio) analyzer.

Techniques: Functional Assay, Inhibition, Activation Assay, Flow Cytometry, Expressing, Incubation, Two Tailed Test, MANN-WHITNEY