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: Journal for Immunotherapy of Cancer

Article Title: Development of therapeutic monoclonal antibodies against DKK1 peptide-HLA-A2 complex to treat human cancers

doi: 10.1136/jitc-2023-008145

Figure Lengend Snippet: Dickkopf-1 -A2 mAbs are therapeutic against established tumors in vivo. (A–D) SCID mice were injected intravenously with luciferase-labeled U266 MM cells, followed by treatment with intraperitoneal injections (every 3 days for a total of six injections) of 200 µg C2 mAb or mIgG. PBS served as a control. Tumor burdens were monitored by (A) in vivo imaging, (B) levels of circulating human M-protein or its light chain, and (C) in vivo bioluminescence plus X-ray imaging. Mouse survival is also shown in (B). (D) Three weeks after the treatment, bone marrow cells and splenocytes were collected for analyzing human CD138 + MM cells using flow cytometry. Summary data is shown. (E–F) SCID mice were injected subcutaneously with Granta 519 lymphoma cells, followed by treatment with intraperitoneal injections (every 3 days for a total of six injections) of 200 µg C2 mAb or mIgG. PBS served as a control. Tumor burdens were measured twice every week by (E) IVIS imaging system and (F) tumor sizes measured by caliper. Mouse survival is shown in (F). (G–H) SCID mice were injected subcutaneously with ARH-77 leukemia cells, followed by treatment with intraperitoneal injections (every 3 days for a total of six injections) of 200 µg C2 mAb or mIgG. PBS served as a control. Tumor burdens were measured twice every week by (G) IVIS imaging system and (H) caliper. Mouse survival is shown in (H). Error bars=SD. *p<0.05; **p<0.01. mAb, monoclonal antibodies; mIgG1, mouse IgG1; MM, multiple myeloma; PBS, phosphate-buffered saline.

Article Snippet: Tumor burdens were evaluated by bioluminescence on the IVIS Spectrum In Vivo Imaging System (PerkinElmer, Waltham, Massachusetts, USA) and the level of human immunoglobulin light chain secreted by human MM cells was determined by ELISA.

Techniques: In Vivo, Injection, Luciferase, Labeling, Control, In Vivo Imaging, Imaging, Flow Cytometry, Saline

Journal: Journal for Immunotherapy of Cancer

Article Title: Development of therapeutic monoclonal antibodies against DKK1 peptide-HLA-A2 complex to treat human cancers

doi: 10.1136/jitc-2023-008145

Figure Lengend Snippet: DKK1-A2 mAbs are therapeutic against established tumors in vivo. (A–C) SCID mice were injected subcutaneously with CFPAC-1 PDAC cells, followed by treatment with intraperitoneal injections (every 3 days for a total of six injections) of 200 µg C2 mAb or mIgG. PBS served as a control. Tumor burdens were measured twice every week by (A) IVIS imaging system and (B) caliper. Mouse survival is shown in (C). (D–F) SCID mice were injected subcutaneously with MDA-MB-231 breast cancer cells, followed by treatment with intraperitoneal injections (every 3 days for a total of six injections) of 200 µg C2, HMB1 or HBM7 mAb or mIgG1. Tumor burdens were measured twice every week by (D) IVIS imaging system and (E) caliper. Mouse survival is shown in (F). Error bars=SD. *p<0.05; **p<0.01. mAb, monoclonal antibodies; mIgG1, mouse IgG1; PBS, phosphate-buffered saline; PDAC, pancreatic ductal adenocarcinoma.

Article Snippet: Tumor burdens were evaluated by bioluminescence on the IVIS Spectrum In Vivo Imaging System (PerkinElmer, Waltham, Massachusetts, USA) and the level of human immunoglobulin light chain secreted by human MM cells was determined by ELISA.

Techniques: In Vivo, Injection, Control, Imaging, Saline

Journal: Journal for Immunotherapy of Cancer

Article Title: Development of therapeutic monoclonal antibodies against DKK1 peptide-HLA-A2 complex to treat human cancers

doi: 10.1136/jitc-2023-008145

Figure Lengend Snippet: In vivo therapeutic effects of Dickkopf-1-A2 mAbs depend on NK cells and are enhanced by lenalidomide. (A–C) SCID mice were xenografted subcutaneously with U266 MM cells followed by treatment with 200 µg mIgG1 or C2 mAb, C2 mAb combined with NK cell depletion by injection of 100 µg specific antibodies (NK depletion), or C2 mAb combined with complement depletion by injection of cobra venom factor (CVF; CP depletion). Tumor burdens were measured twice every week by (A) IVIS imaging system and (B) caliper. Mouse survival is shown in (C). (D–F) SCID mice were challenged subcutaneously with U266 MM cells, followed by treatment with intraperitoneal injections of 200 µg C2 mAb or mIgG1 (every 3 days for a total of six injections), lenalidomide (10 mg/kg, for 5 days and 2 days off for 3 weeks) or lenalidomide plus C2 mAb. Tumor burdens were measured twice every week by (D) IVIS imaging system and (E) caliper. Mouse survival is shown in (F). Representative results of one out of two independent experiments performed are shown. Error bars=SD. *p<0.05, compared with mIgG1 control. mAb, monoclonal antibodies; mIgG1, mouse; NK, natural killer.

Article Snippet: Tumor burdens were evaluated by bioluminescence on the IVIS Spectrum In Vivo Imaging System (PerkinElmer, Waltham, Massachusetts, USA) and the level of human immunoglobulin light chain secreted by human MM cells was determined by ELISA.

Techniques: In Vivo, Injection, Combined Bisulfite Restriction Analysis Assay, Imaging, Control

Journal: ACS applied materials & interfaces

Article Title: Manganese-Loaded pH-Responsive DNA Hydrogels Enable Tg-Guided Thyroid Tumor Targeted Magnetic Resonance Imaging.

doi: 10.1021/acsami.4c19676

Figure Lengend Snippet: Figure 3. In vitro stability and cytotoxicity of M-TDH. (a) Tyndall effect showed good stability of M-TDH in saline with 10% FBS. (b) Cell viability after 24 h of incubation with PBS, TDH, M-TDH, and MnCl2, detected by CCK-8. (c, d) Cell apoptosis rates of DTC cells detected by flow cytometry. (e, f) Live/dead cell staining images and corresponding quantitative comparison of TPC1 and K1 cells after various treatments (scale bar 100 μm). (g, h) Hematological parameters of M-TDH detected by hemolysis experiments and corresponding quantitative measurements.

Article Snippet: The quantification of cellular uptake was obtained by full spectrum analysis flow cytometry (ID7000 Sony, Japan).

Techniques: In Vitro, Saline, Incubation, CCK-8 Assay, Flow Cytometry, Staining, Comparison

Journal: ACS applied materials & interfaces

Article Title: Manganese-Loaded pH-Responsive DNA Hydrogels Enable Tg-Guided Thyroid Tumor Targeted Magnetic Resonance Imaging.

doi: 10.1021/acsami.4c19676

Figure Lengend Snippet: Figure 5. Uptake efficiency and aptamer-mediated combination. (a, b) The uptake efficiency of M-TDH or TDH by K1 and TPC-1 cells detected by flow cytometry. (c) The in situ targeting of the nanohydrogel in frozen thyroid tissue sections. The white box shows the enlarged area, Cy5- TDH shows a red fluorescence signal, and a green fluorescence signal indicates the Tg distribution. (d) Nucleic acid−protein binding assay used to compare the combination between DH or TDH and endogenous Tg protein in the cellular environment.

Article Snippet: The quantification of cellular uptake was obtained by full spectrum analysis flow cytometry (ID7000 Sony, Japan).

Techniques: Flow Cytometry, In Situ, Fluorescence, Protein Binding