stimulation with atp  (MedChemExpress)

Journal: Journal of Nanobiotechnology

Article Title: Fisetin carbon dots alleviate periodontitis by enhancing mitophagy through regulation of sirtuin 3 SUMOylation

doi: 10.1186/s12951-025-03907-9

Figure Lengend Snippet: FIS-CDs alleviated H 2 O 2 -induced oxidative stress, restored mitochondrial function, and promoted mitophagy. ( A ) Assessment of intracellular ROS accumulation via DCFH-DA based fluorescence staining. ( B ) Visualization of cell morphology by staining F-actin with phalloidin. ( C ) Measurement of MDA content in hPDLCs. ( D ) Assessment of T-AOC in hPDLCs. ( E ) Fluorescence images showing the co-localization of FIS-CDs with MitoTracker Green. ( F ) Quantitative analysis of the co-localization between FIS-CDs and mitochondria. ( G ) MitoSOX staining for evaluation of mtROS production. ( H ) Quantitative analysis of MitoSOX signal intensity. (I ) JC-1 staining for assessment of MMP. ( J ) Quantitative analysis of JC-1 signal intensity. ( K–O ) Western blot analysis of mitophagy proteins: ( L ) PINK1, ( M ) Parkin, ( N ) p62, and ( O ) LC3B. ( P ) Quantification of DCFH-DA fluorescence intensity. ( Q ) Quantification of intracellular ATP levels. The data are presented as mean ± SD ( n = 3). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant

Article Snippet: The intracellular concentration of adenosine triphosphate (ATP) was quantified utilizing a commercially available ATP detection kit (MCE, USA).

Techniques: Fluorescence, Staining, Western Blot

Journal: Antioxidants

Article Title: Ferroptosis Enhances T Lymphocyte Infiltration into Glioblastoma Spheroids

doi: 10.3390/antiox14111373

Figure Lengend Snippet: ATP mediates T cell infiltration into glioblastoma spheroids. ( A ) Schematic outline of the experimental setup for the spheroid infiltration assay including ATP degradation by apyrase (Figure created in BioRender. Fuhrmann, D. (2025) https://BioRender.com/8nr6inp ). ( B ) LN229 glioblastoma spheroids were treated with 1 µM RSL3 ± 1 µM liproxstatin-1 (Lip) and apyrase (10 U/mL). ATP release was measured over 24 h and visualized as area under the curve (AUC). ( C ) Leukocytes infiltrating DMSO- and RSL3-treated spheroids, with and without addition of apyrase, were analyzed by flow cytometry after 2 days of co-culture. Data from 15 individual donors. ( D ) Populations of peripheral blood mononuclear cells (PBMCs) infiltrating DMSO- and RSL3-treated spheroids, with and without addition of apyrase, were analyzed by flow cytometry after 2 days of co-culture. PBMC populations were normalized to the respective number of infiltrated leukocytes in each individual sample to generate comparability among the different treatments. Data from 15 individual donors. ( E ) PBMCs were incubated with BAY-1797 (10 µM), a purinergic receptor inhibitor, prior to migration towards living and ferroptotic (1 µM RSL3) glioblastoma cells. Cell migration was analyzed by flow cytometry after 3 h. Data are expressed as mean values ± SEM. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p < 0.0001; p values were calculated using ordinary one-way ANOVA and Tukey’s multiple comparisons test if not stated otherwise.

Article Snippet: ATP receptors were blocked with the P2X4 receptor inhibitor BAY-1797 (HY-130605, MedChemExpress, Princeton, NJ, USA) at 10 μM, which inhibits P2X4 and P2X7 receptors.

Techniques: Flow Cytometry, Co-Culture Assay, Incubation, Migration