Journal: The Journal of Physiological Sciences : JPS

Article Title: Orai1- and STIM1-mediated calcium signaling controls PD-L1 expression and modulates antitumor immunity in oral cancer

doi: 10.1016/j.jphyss.2026.100063

Figure Lengend Snippet: Pharmacological inhibition of SOCE suppresses IFN-γ–induced PD-L1 expression in OSCC cells. (A) HSC-3 cells were pretreated with the SOCE inhibitor Synta66 (1–20 μM) and stimulated with IFN-γ (100 ng/mL, 6 h). PD-L1 and GAPDH protein levels were analyzed by Western blot. PD-L1 is detected as multiple bands, which most likely reflect heterogeneous N-linked glycosylation and maturation states of the protein. (B) PD-L1 expression was examined in cells treated with Synta66 (20 μM) alone under unstimulated conditions. (C) Cells were pretreated with Synta66 (20 μM, 3 h), followed by IFN-γ (100 ng/mL, 3 h). PD-L1 mRNA was assessed by RT-qPCR. (D) Cells were pretreated with Synta66 (20 μM, 3 h), followed by IFN-γ (100 ng/mL, 6 h). PD-L1 protein levels were analyzed by Western blot. Quantitative data are shown as mean ± SEM from four independent biological replicates (n = 4). Exact p-values or significance levels (p < 0.001 or ns) are indicated in the figure. Representative Western blot images are shown.

Article Snippet: The SOCE channel inhibitor Synta66, which targets Orai1 forming the pore of the CRAC channel, was purchased from MedChemExpress (Monmouth Junction, NJ, USA), respectively.

Techniques: Inhibition, Expressing, Western Blot, Glycoproteomics, Quantitative RT-PCR

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