ionomycin  (Tocris)

Journal: bioRxiv

Article Title: Oleuropein stimulates peripheral serotonin secretion via voltage-dependent Ca 2+ channels and acutely regulates central function

doi: 10.64898/2026.07.15.738825

Figure Lengend Snippet: Effects of oleuropein on 5-HT secretion from enterochromaffin-like cells and Ca 2+ influx. ( A ) Structure of oleuropein. ( B ) Dose-dependent effects of oleuropein on 5-HT secretion. QGP-1 cells were incubated with HBSS containing different concentrations of oleuropein and the supernatant was subjected to HPLC analysis. ( C ) Dose-dependent effects of oleuropein on cell viability. QGP-1 cells were incubated with HBSS containing different concentrations of oleuropein for 60 min and subjected to the WST-8 assay. ( D ) Dose-dependent effects of oleuropein on cytosolic Ca 2+ levels. Fluo-8 AM was internalized into QGP-1 cells, and oleuropein was added. The fluorescence intensity was then measured over time for 5 min. Ionomycin was used as a positive control. ( E ) Quantification of changes in the Fluo-8 fluorescence intensity. The average change in fluorescence intensity from 0 min after oleuropein addition to each time point was expressed as Δ[Ca 2+ ]i (ΔF/F0). ( F ) Effects of cell-membrane-impermeable Ca 2+ chelator on 5-HT secretion by oleuropein. QGP-1 cells were incubated with HBSS containing 100 µM oleuropein for 5 min with or without BAPTA, and the supernatant was subjected to HPLC analysis. ( G ) Effects of cell-membrane-permeable Ca 2+ chelator on 5-HT secretion by oleuropein. QGP-1 cells were incubated with HBSS containing 100 µM oleuropein for 5 min with or without BAPTA-AM and the supernatant was subjected to HPLC analysis. Bars represent the mean ± SEM, and dots indicate individual data points ( n = 3). ** p < 0.01, *** p < 0.001, and **** p < 0.0001, one-way ANOVA, Tukey’s multiple comparisons test. ns denotes not significant.

Article Snippet: Then, oleuropein, hydroxytyrosol or ionomycin (Alomone Labs) was added as the positive control, and the fluorescence intensity (Ex/Em = 492/530 nm) was measured every 15 sec for 300 sec.

Techniques: Incubation, Analysis, Fluorescence, Positive Control, Membrane

Journal: bioRxiv

Article Title: Oleuropein stimulates peripheral serotonin secretion via voltage-dependent Ca 2+ channels and acutely regulates central function

doi: 10.64898/2026.07.15.738825

Figure Lengend Snippet: Effects of hydroxytyrosol on 5-HT secretion from enterochromaffin-like cells and Ca 2+ influx. ( A ) Structure of hydroxytyrosol. ( B ) Dose-dependent effects of hydroxytyrosol on 5-HT secretion. QGP-1 cells were incubated with HBSS containing different concentrations of hydroxytyrosol, and the supernatant was subjected to HPLC analysis. ( C ) Dose-dependent effects of hydroxytyrosol on cell viability. QGP-1 cells were incubated with HBSS containing different concentrations of hydroxytyrosol for 60 min and subjected to the WST-8 assay. ( D ) Dose-dependent effects of hydroxytyrosol on cytosolic Ca 2+ levels. Fluo-8 AM was internalized into QGP-1 cells, and hydroxytyrosol was added. The fluorescence intensity was then measured over time for 5 min. Ionomycin was used as a positive control. The average change in fluorescence intensity from 0 min after hydroxytyrosol addition to each time point was expressed as Δ[Ca 2+ ]i (ΔF/F0). ( E ) Effects of T-type voltage-dependent Ca 2+ channel inhibitor on 5-HT secretion by hydroxytyrosol. QGP-1 cells were incubated with HBSS containing 100 µM hydroxytyrosol for 5 min with or without NNC 55-0396. ( F ) Effects of T-type voltage-dependent Ca 2+ channel inhibitor on Ca 2+ influx induced by hydroxytyrosol. Fluo-8 AM was internalized into QGP-1 cells and hydroxytyrosol was added with or without NNC 55-0396. The fluorescence intensity was then measured over time for 5 min, and the average change in fluorescence intensity from 0 min after hydroxytyrosol addition to each time point was expressed as Δ[Ca 2+ ]i (ΔF/F0). ( G ) Effects of L-type voltage-dependent Ca 2+ channel inhibitor on 5-HT secretion by hydroxytyrosol. QGP-1 cells were incubated with HBSS containing 100 µM hydroxytyrosol for 5 min, with or without nifedipine. ( H ) Effects of L-type voltage-dependent Ca 2+ channel inhibitor on Ca 2+ influx induced by hydroxytyrosol. Fluo-8 AM was internalized into QGP-1 cells, and hydroxytyrosol was added with or without nifedipine. The fluorescence intensity was then measured over time for 5 min and the average change in fluorescence intensity from 0 min after hydroxytyrosol addition to each time point was expressed as Δ[Ca 2+ ]i (ΔF/F0). Bars represent the mean ± SEM, and dots indicate individual data points ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, one-way ANOVA, Tukey’s multiple comparisons test. ns denotes not significant.

Article Snippet: Then, oleuropein, hydroxytyrosol or ionomycin (Alomone Labs) was added as the positive control, and the fluorescence intensity (Ex/Em = 492/530 nm) was measured every 15 sec for 300 sec.

Techniques: Incubation, Analysis, Fluorescence, Positive Control

Journal: Leukemia

Article Title: Leukemia-driven expansion of type 3 innate lymphoid cell facilitates a pro-tumoral microenvironment in acute myeloid leukemia

doi: 10.1038/s41375-025-02829-7

Figure Lengend Snippet: A Summary data of Ki67 frequency from ILC3 from congenic Ly5.1 transplanted murine PTD/ITD (PTD partial tandem duplication, ITD internal tandem duplication) AML model versus WT, n = 10, ** p < 0.01. Error bars represent ± SEM. B Murine ILC3-derived cytokines frequency of ILC3 determined by flow cytometry from leukemic bone marrow relative to non-leukemic controls post-stimulation with PMA/Ionomycin, IL-1β, IL-23 and IL-7, n = 9–10, ** p < 0.01, **** p < 0.0001. Error bars represent ± SEM. C Flow cytometry data of expression of ILC3-derived cytokines following PMA/Ionomycin, IL-1β, IL-23 and IL-7 stimulation of ILC3s cultured in vitro with the AML cell line (U937) relative to ILC3 alone controls, n = 10, * p < 0.05. Error bars represent ± SEM. D Flow cytometry data of expression of ILC3-derived cytokines in the bone marrow (BM) of untreated AML patients relative to healthy marrow, post-stimulation of PMA/Ionomycin, IL-1β, IL-23 and IL-7, n = 8 Normal BM, n = 7 AML BM, ** p < 0.01. Error bars represent ± SEM.

Article Snippet: ILC3s or bulk single cell suspensions were harvested and evaluated at baseline or stimulated with PMA/Ionomycin (eBioscience) and 50 ng/mL each of IL-23, IL-1β, and IL-7, for 2 h followed by the addition of Brefeldin A (10 ug/ml) for an additional 10–12 h. Cells were then harvested and phenotyped as previously described [ ].

Techniques: Derivative Assay, Flow Cytometry, Expressing, Cell Culture, In Vitro