Journal: Cancers

Article Title: NIR Imaging of the Integrin-Rich Head and Neck Squamous Cell Carcinoma Using Ternary Copper Indium Selenide/Zinc Sulfide-Based Quantum Dots

doi: 10.3390/cancers12123727

Figure Lengend Snippet: Expression of integrins in FaDu and MeWo cells. ( a ) Western blot of β 6 subunit expression in FaDu and MeWo cells and its quantification analysis. Data represent averages ± SD (n = 5; * p < 0.05, using the one-sample t-test (µ = 1)). ( b , c ) Typical fluorescence microscopy images of ( b ) F5 and ( c ) F5M5 spheroids cryosections, stained with antibody against α v β 6 integrins and DAPI. MeWo cells were pre-stained with PKH67 green membrane dye. Scale bar—100 μm.

Article Snippet: As CAFs, we used MeWo cells (ATCC HTB-65), granular fibroblasts derived from human melanoma [ ], cultured in Minimal Essential Medium (MEM, Sigma-Aldrich, Saint-Quentin Fallavier, France), supplemented with 9% ( vol / vol ) of FBS and 1% ( vol / vol ) 0.1 M sodium pyruvate (Sigma-Aldrich, Saint-Quentin Fallavier, France).

Techniques: Expressing, Western Blot, Fluorescence, Microscopy, Staining, Membrane

Journal: Cancers

Article Title: NIR Imaging of the Integrin-Rich Head and Neck Squamous Cell Carcinoma Using Ternary Copper Indium Selenide/Zinc Sulfide-Based Quantum Dots

doi: 10.3390/cancers12123727

Figure Lengend Snippet: Uptake of QDs in 2D monolayer FaDu and MeWo cells. Typical cytometry distribution histograms of ( a ) FaDu and ( b ) MeWo monolayer cells 3 h post-incubation with 50 nM of QDs. ( c ) Mean fluorescence intensity (MFI) of FaDu (cyan bars) and MeWo (orange bars) cells 3 h post-incubation with QDs (n = 6–7; *** p < 0.001 compared to autofluorescence, using ANOVA; †† p < 0.01, using two-sample t-test).

Article Snippet: As CAFs, we used MeWo cells (ATCC HTB-65), granular fibroblasts derived from human melanoma [ ], cultured in Minimal Essential Medium (MEM, Sigma-Aldrich, Saint-Quentin Fallavier, France), supplemented with 9% ( vol / vol ) of FBS and 1% ( vol / vol ) 0.1 M sodium pyruvate (Sigma-Aldrich, Saint-Quentin Fallavier, France).

Techniques: Cytometry, Incubation, Fluorescence

Journal: Cancers

Article Title: NIR Imaging of the Integrin-Rich Head and Neck Squamous Cell Carcinoma Using Ternary Copper Indium Selenide/Zinc Sulfide-Based Quantum Dots

doi: 10.3390/cancers12123727

Figure Lengend Snippet: Uptake of QDs in 3D spheroids. ( a ) Flow cytometry histograms of cells from monoculture (F5) spheroids exposed for 3 h to 50 nM QD-SPP (blue) and A20-QDs (red). ( b ) The MFI and ( c ) the number of labeled FaDu cells in F5 spheroids exposed to various concentrations of QD-SPP (blue) and QD-A20 (red). MFI of ( d ) FaDu; ( e ) MeWo cells and ( f ) total number of labeled cells in F5M5 spheroids in the function of QD concentration. Typical flow cytometry histograms of ( g ) FaDu and ( h ) MeWo cells from F5M5 spheroids exposed for 3 h to 50 nM QD-SPP (blue) and A20-QDs (red). ( i ) The fraction of FaDu (cyan) and MeWo (orange) cells from F5M5 spheroids in the function of QD concentration. Data represent mean ± SD (n = 4–7; ** p < 0.01; *** p < 0.001 compared to autofluorescence, using ANOVA; † p < 0.05, †† p < 0.01 and ††† p < 0.001, using two-sample t-test).

Article Snippet: As CAFs, we used MeWo cells (ATCC HTB-65), granular fibroblasts derived from human melanoma [ ], cultured in Minimal Essential Medium (MEM, Sigma-Aldrich, Saint-Quentin Fallavier, France), supplemented with 9% ( vol / vol ) of FBS and 1% ( vol / vol ) 0.1 M sodium pyruvate (Sigma-Aldrich, Saint-Quentin Fallavier, France).

Techniques: Flow Cytometry, Labeling, Concentration Assay

Journal: Cancers

Article Title: NIR Imaging of the Integrin-Rich Head and Neck Squamous Cell Carcinoma Using Ternary Copper Indium Selenide/Zinc Sulfide-Based Quantum Dots

doi: 10.3390/cancers12123727

Figure Lengend Snippet: 3D Fluorescence imaging of QD-loaded head and neck squamous cell carcinoma (HNSCC) spheroids. 3D confocal microscopy of QD-A20 (red color, λ em = 730–800 nm) distribution in different optical sections of ( a ) F5 and ( b ) F5M5 spheroids, incubated for 3 h with 50 nM of QDs. MeWo cells were pre-stained with PKH67 membrane dye (green color, λ em = 530–600 nm). The orange line displays the contour of spheroid. The mean pixel intensity of the central region (blue circles) of ( c ) F5 and ( d ) F5M5 spheroids 3 h post-incubation with 50 nM of QDs as a function of the depth (z) of optical section. Scale bar = 100 µm. Data represent mean ± SD (n = 3–7; ** p < 0.01; *** p < 0.001 compared to autofluorescence, using ANOVA).

Article Snippet: As CAFs, we used MeWo cells (ATCC HTB-65), granular fibroblasts derived from human melanoma [ ], cultured in Minimal Essential Medium (MEM, Sigma-Aldrich, Saint-Quentin Fallavier, France), supplemented with 9% ( vol / vol ) of FBS and 1% ( vol / vol ) 0.1 M sodium pyruvate (Sigma-Aldrich, Saint-Quentin Fallavier, France).

Techniques: Fluorescence, Imaging, Confocal Microscopy, Incubation, Staining, Membrane

Journal: International journal of molecular medicine

Article Title: Cynandione A inhibits lipopolysaccharide-induced cell adhesion via suppression of the protein expression of VCAM‑1 in human endothelial cells.

doi: 10.3892/ijmm.2018.3376

Figure Lengend Snippet: Figure 3. Effects of CA on the phosphorylation of mitogen‑activated protein kinase/IKK/IκB and analysis of the translocation of p65 into the nucleus of LPS‑activated HUVECs. (A) HUVECs were pretreated with the indicated signal inhibitor, AG490 (10 µM), U0126 (10 µM), SB203580 (10 µM) or PDTC (10 µM) for 1 h, and stimulated with LPS with or without CA for 24 h. mRNA and protein expression levels of VCAM‑1 were detected using reverse transcription‑quantitative polymerase chain reaction and western blot analyses, respectively. (B) HUVECs were starved for 6 h and then pre‑treated with CA for 1 h, followed by activation by LPS for 30 min. Phosphorylation patterns were analyzed using western blot analysis. (C) HUVECs were starved for 6 h and then pre‑treated with CA for 1 h, followed by activation by LPS for 30 min. The activation levels of p‑IKK, p‑IκB‑α, IKK and IκB‑α were detected using western blot analysis. (D) HUVECs were starved for 6 h and then pre‑treated with CA for 1 h, followed by activation with LPS for 2 h. Proteins in the nuclear extract fraction were prepared and NF‑κB p65 subunit translocation levels were analyzed using western blot analysis. **P<0.01, compared with the negative control; *P<0.05, compared with treatment with LPS alone. HUVECs, human umbilical vascular endothelial cells; CA, cynandione A; LPS, lipopolysac- charide; VCAM‑1, vascular adhesion molecule‑1; NF‑κB, nuclear factor‑κB; IκB, inhibitor of NF‑κB; IKK, IκB kinase; JNK, c‑Jun N‑terminal kinase; ERK, extracellular signal‑regulated kinase; p‑, phosphorylated.

Article Snippet: Antibodies against p-IKK α/β (87, 85 kda, 1:1,000; cat no. #2697) p-IκB-α (40 kda, 1:2,000; cat no. #9246), anti-p65 (65 kda, 1:1,000; cat no. #8242) and all non-phosphorylated antibodies were prepared and acquired from cell Signaling Technology, Inc. (Beverly, MA, USA).

Techniques: Phospho-proteomics, Translocation Assay, Expressing, Polymerase Chain Reaction, Western Blot, Activation Assay, Negative Control

Journal: International journal of molecular medicine

Article Title: Cynandione A inhibits lipopolysaccharide-induced cell adhesion via suppression of the protein expression of VCAM‑1 in human endothelial cells.

doi: 10.3892/ijmm.2018.3376

Figure Lengend Snippet: Figure 4. Effects of CA on the transcriptional activity of NF‑κB and luciferase reporter genes in transiently transfected HUVECs. (A) pVCAM‑1/Luc trans- fected HUVECs were cultured for 24 h, and treated with indicated concentrations of CA and 1 µg/ml LPS for 24 h. The values for relative luciferase intensity are shown as the mean ± standard deviation of three independent experiments (n=3). **P<0.01, compared with the mock transfectant; **P<0.01, compared with treatment with LPS alone. (B) pNF‑κB/Luc transfected HUVECs were cultured for 24 h, and treated with indicated concentrations of CA and 1 µg/ml LPS for 24 h. The values for relative luciferase intensity are shown as the mean ± standard deviation from three independent experiments (n=3). *P<0.05, compared with the mock transfectant; *P<0.05, compared with treatment with LPS alone. (C) Nuclear translocation of NF‑κB p65 subunit was analyzed following CA (40 µM) and PDTC (10 µM) treatment for 1 h. Cells were activated for 2 h with LPS and then observed via fluorescence microscopy (magnification, x100). HUVECs, human umbilical vascular endothelial cells; CA, cynandione A; LPS, lipopolysaccharide; NF‑κB, nuclear factor κB; VCAM‑1, vascular adhesion molecule‑1; Luc, luciferase.

Article Snippet: Antibodies against p-IKK α/β (87, 85 kda, 1:1,000; cat no. #2697) p-IκB-α (40 kda, 1:2,000; cat no. #9246), anti-p65 (65 kda, 1:1,000; cat no. #8242) and all non-phosphorylated antibodies were prepared and acquired from cell Signaling Technology, Inc. (Beverly, MA, USA).

Techniques: Activity Assay, Luciferase, Transfection, Cell Culture, Standard Deviation, Translocation Assay, Fluorescence, Microscopy

Journal: Redox Biology

Article Title: PATJ regulates cell stress responses and vascular remodeling post-stroke

doi: 10.1016/j.redox.2025.103709

Figure Lengend Snippet: YAP1 expression and localization in PATJ-modified HEK293 cells and ischemic stroke.(A) Western blot analysis of YAP1 protein levels in WT and PATJ KO cells, with quantification showing a significant increase in YAP1 protein levels in KO cells (n = 4 for each group). (B) Immunofluorescence staining of YAP1 (green) DAPI staining of nuclei (blue). Yellow arrows indicate cells with predominantly cytoplasmic YAP1 localization, red arrowheads indicate predominantly nuclear YAP1 localization, and white arrowheads indicate cells with YAP1 distributed in both compartments. Right panels show quantitative analysis of YAP1 subcellular distribution patterns, presented as the percentage of cells with YAP1 localization predominantly in the nucleus, cytoplasm, or both compartments (n = 4 independent experiments with >50 cells/group). (C) Confocal microscopy with 3D reconstruction of YAP1 subcellular distribution in WT and PATJ KO cells. Right panel shows quantification of the YAP1 nuclear-to-cytoplasmic intensity ratio (n = 4 independent experiments with 50 cells/group). (D) Scratch-wound assay of WT and KO monolayers treated with verteporfin, captured at 0, 24, and 48 h post-scratch. Dashed lines denote wound edge (n = 4 independent experiments with 3 technical replicates/group). (E) Cell survival following 48 h exposure to rotenone with or without verteporfin (n = 3 independent experiments with 3 technical replicates/group). (F) Representative images of endothelial cell marker CD31 (red) and YAP1 (green) in brain tissue from sham and 28 days post-middle cerebral artery occlusion (MCAO) mice, with DAPI staining of nuclei (blue) and merged images highlighting co-localization. n = 4 mice per group; 3 non-consecutive sections per mouse with 5 randomly selected peri-infarct fields analyzed per section. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: For YAP1/CD31/DAPI triple staining, sections were incubated overnight at 4 °C with rat anti-mouse CD31 (1:200, BD Pharmingen) and rabbit polyclonal YAP1 antibody (1:200, Proteintech).

Techniques: Expressing, Modification, Western Blot, Immunofluorescence, Staining, Confocal Microscopy, Scratch Wound Assay Assay, Marker

Journal: Redox Biology

Article Title: PATJ regulates cell stress responses and vascular remodeling post-stroke

doi: 10.1016/j.redox.2025.103709

Figure Lengend Snippet: Temporal expression of PATJ in endothelial cells post-tMCAO and colocalization with YAP1. (A)Schematic representation of the peri-ischemic region observed in the tMCAO mouse model. (B) Representative immunofluorescence images showing double staining of Patj (red) and CD31 (green) in brain sections from sham and tMCAO-treated mice at days 1, 3, 7, 14, and 28 post-stroke. (C) Quantitative analysis of Patj/CD31 double positive cells per mm 2 in the peri-ischemic region over time (D) Quantitative analysis of Patj positive cells in the peri-ischemic area over time. (E) Quantitative analysis of CD31 positive signal dynamics in the peri-ischemic zone. n = 4 mice per group; 3 non-consecutive sections per mouse with 5 randomly selected peri-infarct fields analyzed per section. ∗∗p < 0.01, ∗∗∗∗p < 0.0001. (F) Triple immunofluorescence staining showing CD31 (red), YAP1 (green), and PATJ (blue) in brain sections from sham and 7 days post-tMCAO mice. The merged images demonstrate colocalization of all three markers, with white arrowheads indicating triple-positive regions in the vascular structures. (G) Quantification shows the percentage of CD31+/YAP1+/PATJ + triple-positive area in sham versus tMCAO 7d tissues. n = 4 mice per group; 3 non-consecutive sections per mouse with 5 randomly selected peri-infarct fields analyzed per section. ∗∗∗∗p < 0.0001.

Article Snippet: For YAP1/CD31/DAPI triple staining, sections were incubated overnight at 4 °C with rat anti-mouse CD31 (1:200, BD Pharmingen) and rabbit polyclonal YAP1 antibody (1:200, Proteintech).

Techniques: Expressing, Immunofluorescence, Double Staining, Staining