Journal: Cell death & disease

Article Title: Circular RNA TFRC/SCD1 mRNA interaction regulates ferroptosis and metastasis in gastric cancer.

doi: 10.1038/s41419-025-07759-x

Figure Lengend Snippet: Fig. 4 CircTFRC knockdown promoted the ferroptosis of GC cells. A, B Propidium iodide (PI) staining showing the cell death rates of control (si-NC) and circTFRC knockdown (si-circTFRC) AGS and HGC-27 cells in the absence or presence of ferrostatin-1 (0.75 μM, 16 h). C, D Flow cytometry showing the lipid ROS levels (stained with C11 BODIPY 581/591) in control (si-NC) and circTFRC knockdown (si-circTFRC) AGS and HGC-27 cells in the absence or presence of ferrostatin-1 (0.75 μM, 16 h). ROS reactive oxygen species. E, F ELISA assays showing the relative MDA levels in control (si-NC) and circTFRC knockdown (si-circTFRC) AGS and HGC-27 cells in the absence or presence of ferrostatin-1 (0.75 μM, 16 h). MDA malondialdehyde. G, H Plate colony formation assay assessing colony formation in AGS cells under control conditions (sh-NC) or with circTFRC knockdown (sh-circTFRC) in the absence or presence of ferrostatin-1 (0.25 μM, 72 h). I, J Transwell assay evaluating the migration ability of AGS cells under control conditions (sh-NC) or with circTFRC knockdown (sh-circTFRC) in the absence or presence of ferrostatin-1 (0.75 μM, 16 h). K PI staining showing the cell death rates of control (sh-NC) and circTFRC knockdown (sh-circTFRC) AGS cells following treatment with RSL3 (1 μM) in the absence or presence of ferrostatin-1 (0.75 μM) for 16 h. L Flow cytometry showing the lipid ROS levels (stained with C11 BODIPY 581/591) in control (sh-NC) and circTFRC knockdown (sh-circTFRC) AGS cells following treatment with RSL3 (1 μM) in the absence or presence of ferrostatin-1 (0.75 μM) for 16 h. M Confocal microscopy showing the lipid ROS levels in control (sh-NC) and circTFRC knockdown (sh-circTFRC) AGS cells following treatment with RSL3 (1 μM) in the absence or presence of ferrostatin-1 (0.75 μM) for 16 h. Lipid ROS are stained with C11 BODIPY 581/591 probe (green). Nuclei are counterstained with DAPI (blue). Scale bar, 10 µm. Data are presented as mean ± SD. P-values were calculated using a two-tailed one-way ANOVA (A–F) or unpaired Student’s t test (H, J–L); **P < 0.01, ***P < 0.001, ****P < 0.0001. See also Fig. S4.

Article Snippet: Quantitative analysis of PI-positive populations was subsequently conducted using the Cytek® Northern LightsTM-CLC (NL-CLC) full spectrum flow cytometry system (Cytek® Biosciences, USA), with a minimum of 10,000 cells analyzed per condition.

Techniques: Knockdown, Staining, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Colony Assay, Transwell Assay, Migration, Confocal Microscopy, Two Tailed Test

Journal: Cell death & disease

Article Title: Circular RNA TFRC/SCD1 mRNA interaction regulates ferroptosis and metastasis in gastric cancer.

doi: 10.1038/s41419-025-07759-x

Figure Lengend Snippet: Fig. 6 CircTFRC exerted an oncogenic effect via SCD1 in GC cells. A CCK-8 assay showing the proliferation of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection at indicated time points (0, 24, 48, and 72 h). B, C Plate colony formation assay assessing colony formation in AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection over 10 days. D, E Transwell assay showing the migration of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection over 24 h. Scale bar, 100 µm. F PI staining showing the cell death rates of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection. G Flow cytometry showing the lipid ROS levels (stained with C11 BODIPY 581/591) of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection. ROS reactive oxygen species. H ELISA assays showing the relative MDA levels of AGS cells under control conditions (sh-NC), or upon circTFRC knockdown (sh-circTFRC) or sh-circTFRC + SCD1 vector cotransfection. MAD malondialdehyde. The data are shown as the mean ± SD. The P values were determined by a two-tailed unpaired Student’s t test (C, E–H) or two-way ANOVA (A); **P < 0.01, ***P < 0.001, ****P < 0.0001. See also Fig. S7.

Article Snippet: Quantitative analysis of PI-positive populations was subsequently conducted using the Cytek® Northern LightsTM-CLC (NL-CLC) full spectrum flow cytometry system (Cytek® Biosciences, USA), with a minimum of 10,000 cells analyzed per condition.

Techniques: CCK-8 Assay, Control, Knockdown, Plasmid Preparation, Cotransfection, Colony Assay, Transwell Assay, Migration, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Cell Reports Methods

Article Title: Application of spectral flow cytometry for comprehensive detection of immune metabolism in patient-derived microsamples

doi: 10.1016/j.crmeth.2026.101330

Figure Lengend Snippet: Assessing spectral resolvability of metabolic probes for multiparametric cytometry (A) Experimental workflow for evaluating probe co-detection. (B and D) Representative density plots demonstrating spectral overlap for pairwise probe combinations within the (B) FITC and (D) PE channel. (C and E) Complementary validation matrices showing computational spectral similarity scores from FluoroFinder (lower left quadrant) versus empirical resolvability determined by unmixing (upper right quadrant). Gray indicates non-resolvable pairs; colored tiles indicate resolvable pairs (C, green; E, red).

Article Snippet: Cells were washed and then acquired by full spectrum flow cytometry (SONY ID7000).

Techniques: Cytometry, Biomarker Discovery

Journal: Cell Reports Methods

Article Title: Application of spectral flow cytometry for comprehensive detection of immune metabolism in patient-derived microsamples

doi: 10.1016/j.crmeth.2026.101330

Figure Lengend Snippet: Validation framework for spectral co-resolvability of metabolic probes and fluorophore conjugates (A) Experimental workflow for co-detection assessment. (B, D, and F) Representative flow cytometry density plots demonstrating pairwise mixing between metabolic probes and fluorophores in the (B) FITC/AF488, (D) PE, and (F) APC/AF647 channel. (C, E, and G) Validation matrices per channel: upper, computational spectral similarity by FluoroFinder; lower, empirically determined resolvability. Gray tiles indicate non-resolvable pairs; colored tiles confirm resolvable pairs. (C) FITC/AF488, green; (E) PE, red; and (G) APC/AF647, blue.

Article Snippet: Cells were washed and then acquired by full spectrum flow cytometry (SONY ID7000).

Techniques: Biomarker Discovery, Flow Cytometry

Journal: Cell Reports Methods

Article Title: Application of spectral flow cytometry for comprehensive detection of immune metabolism in patient-derived microsamples

doi: 10.1016/j.crmeth.2026.101330

Figure Lengend Snippet: Validation of three metabolic probes for simultaneous assessment of mitochondrial activity and oxidative stress (A) Representative flow cytometry density plots demonstrating spectral resolution of co-stained probes. (B) Validation matrix: lower left quadrant, computational spectral similarity by FluoroFinder; upper right quadrant, empirical resolvability determination (red: resolvable pairs). (C) Mean fluorescence intensity (MFI; mean ± SD) of individual probes in control versus rotenone/antimycin A (Rot/AA)-treated groups ( n = 6). (D) Correlation analysis of MFI between single-stain and multiplexed conditions across probes (Spearman’s r). (E) MFI of probes (mean ± SD) in control versus EZH2-knockdown (EZH2-sh) groups under single and multiplexed staining. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.

Article Snippet: Cells were washed and then acquired by full spectrum flow cytometry (SONY ID7000).

Techniques: Biomarker Discovery, Activity Assay, Flow Cytometry, Staining, Fluorescence, Control, Knockdown