Journal: Nature Communications
Article Title: MASTER-NAADP: a membrane permeable precursor of the Ca 2+ mobilizing second messenger NAADP
doi: 10.1038/s41467-024-52024-y
Figure Lengend Snippet: Global Ca 2+ signaling was analyzed in Jurkat WT T cells ( A ) and Jurkat Hn1l/Jpt2 -/- T cells ( B ), loaded with Fura2-AM. MASTER-NAADP (100 nM), MASTER-NADP (100 nM), or NAADP-AM (100 nM) were added as indicated. Experiments were carried out at 37 °C and SOCE was blocked by pre-incubation of 50 µM Synta66 for 5 min prior to imaging. Number of cells indicated in figure. Aggregated data are mean amplitude per peak ( C ), mean number of Ca 2+ peaks ( D ), percentage of responding cells ( E ), and calculation of the mean responsiveness (number of peaks * amplitude; F presented as mean ± SEM; Jurkat WT MASTER-NAADP, n = 240 cells; Jurkat WT MASTER-NADP, n = 120 cells; Jurkat WT NAADP-AM, n = 91 cells; Jurkat Hn1l/Jpt2 -/- MASTER-NAADP, n = 181 cells; Jurkat Hn1l/Jpt2 -/- MASTER-NADP, n = 113 cells; Jurkat Hn1l/Jpt2 -/- NAADP-AM, n = 110 cells. Nonparametric Kruskal-Wallis test and Dunn’s correction for multiple testing * P < 0.05; **** P < 0.0001. Source data and exact p values are provided as a Source Data file.
Article Snippet: Additionally, Jurkat T cells were incubated with 50 µM Synta66 (Merck) for 5 min prior to imaging to block SOCE through inhibition of Orai1.
Techniques: Incubation, Imaging