Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the HLA-E–NKG2A axis in combination with MS-275 enhances NK cell-based immunotherapy against DMG

doi: 10.1186/s13046-025-03390-y

Figure Lengend Snippet: MS-275 enhances NK cell-mediated cytotoxicity against DMG cells. A - B Luciferase-engineered TT150630 and TT190326 cell lines were treated with DMSO or 1µM MS-275 for 2 days and cocultured with NK-92MI cells at the indicated E:T ratios. After 4 h of incubation, bioluminescence was measured, and NK cell-induced cytotoxicity (%) was calculated and plotted. C - F DMG cells were treated with MS-275 (1µM) for 2 days, stained with calcein AM, and cocultured with NK-92MI cells in 96-well plates at a 1:1 ratio (TT190326, TT150630 and TT170720) or a 5:1 ratio (TT150728). After incubation for 4 h, fluorescence images were captured using an inverted microscope. Images of DMG cell lines showing a reduction in fluorescence (indicating cell death) are presented (left). Calcein AM-stained DMG cells without NK-92MI cells served as controls. The percentage (%) of cells undergoing NK cell-induced cytotoxicity was calculated and plotted (right). Statistical significance was assessed via Student’s t test, with * p < 0.05 and ** p < 0.01. Scale bars, 500 μm

Article Snippet: To stain DMG cells, 1 μM of calcein AM (MCE, Cat# HY-D0041) was added to the medium (1 × 10 6 cells/ml) and the cells were incubated for 60 min at room temperature.

Techniques: Luciferase, Incubation, Staining, Fluorescence, Inverted Microscopy

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the HLA-E–NKG2A axis in combination with MS-275 enhances NK cell-based immunotherapy against DMG

doi: 10.1186/s13046-025-03390-y

Figure Lengend Snippet: MS-275 enhances the efficacy of immunotherapies targeting the NKG2A–HLA-E axis in vitro. A - B Luciferase-engineered TT150630 and TT190326 cell lines were treated with DMSO or 1µM MS-275 for 2 days and cocultured with NK-92MI cells for 4 h at a 1:1 ratio. The NK-92MI cells were preincubated with control IgG (50 μg/ml), tiragolumab (50 μg/ml) or monalizumab (50 μg/ml) for 1 h at 37 °C before being cocultured with the tumor cells and the NK-92MI cells. The percentage (%) of cells undergoing NK cell-induced cytotoxicity was calculated and plotted. Statistical significance was assessed via one-way ANOVA, with * p < 0.05 and ** p < 0.01. C - D TT190326 and TT150630 cells were treated with MS-275 (1µM) for 2 days, stained with calcein AM, and cocultured with NK-92MI cells for 4 h in 96-well plates at a 1:1 ratio. The NK-92MI cells were preincubated with control IgG (50 μg/ml), tiragolumab (50 μg/ml) or monalizumab (50 μg/ml) for 1 h at 37 °C before being cocultured with the tumor cells and the NK-92MI cells. Fluorescence images were captured using an inverted microscope (left). The percentage (%) of cells undergoing NK cell-induced cytotoxicity was calculated and plotted (right). Statistical significance was assessed via one-way ANOVA, with * p < 0.05 and ** p < 0.01. Scale bars, 500 μm.

Article Snippet: To stain DMG cells, 1 μM of calcein AM (MCE, Cat# HY-D0041) was added to the medium (1 × 10 6 cells/ml) and the cells were incubated for 60 min at room temperature.

Techniques: In Vitro, Luciferase, Control, Staining, Fluorescence, Inverted Microscopy

Journal: Stem Cell Research & Therapy

Article Title: Interleukin-1β induces CXCR3-mediated chemotaxis to promote umbilical cord mesenchymal stem cell transendothelial migration

doi: 10.1186/s13287-018-1032-9

Figure Lengend Snippet: Effects of IL-1β in morphology and interaction between HUVECs and MSCs. Labeled MSCs with CellTracker™ Orange were seeded on HUVECs stained with Calcein AM and co-cultivated for 30 to 240 min. After a period of 60 min, MSCs attached to HUVECs and the morphology were still spherical but developed form of cytoplasmic offshoot. a After 60 min, MSC became flattened and adhered to HUVEC monolayer. IL-1β promoted adhesion (left, 60 min) and transendothelial migration abilities (right, 240 min) of MSCs. b After 180 and 240 min, MSCs extended long plasmic filopodia and integrated into the HUVEC monolayer. Orthogonal projections illustrate that MSCs inserted into HUVEC monolayer (left, 180 min) and formation of filopodia caused transendothelial migration (right, 240 min). Arrows indicate MSC migration through the HUVEC. Horizontal bar: XZ plane of confocal image stack; vertical bar: YZ plane of confocal image stack. Scale bar = 10 μm

Article Snippet: HUVEC monolayers were stained with 8 μM Calcein AM (Tocris, UK) in serum-free medium for 30 min at 37 °C, then gently washed three times with PBS.

Techniques: Labeling, Staining, Migration