Fig. 1: HEI3090 enhances ATP-induced receptor channel activity. | Nature Communications

Fig. 1: HEI3090 enhances ATP-induced receptor channel activity.

From: A small-molecule P2RX7 activator promotes anti-tumor immune responses and sensitizes lung tumor to immunotherapy

Fig. 1

a Representation of HEI3090’s synthesis steps. b Modulation of ATP-induced intracellular Ca2+ variation (F1/F0) in HEK293T-mP2RX7 cells (C57Bl/6 origin). After ten baseline cycles, ATP (333 µm) and HEI3090 (250 nM) were injected. Error bars are mean ± SEM (n = 3 independent experiments, 6 replicates). c Average Fluo-4-AM fluorescence intensities in HEK293T mP2RX7 or control HEK pcDNA6 measured 315 s after stimulation with ATP (333 µM) and HEI3090 at concentrations of 25, 250, and 2.5 µM, as indicated in the color code. Data are presented as scatter dot plots ± SEM (n = 3 independent experiments and 6 replicates, two-tailed Mann–Whitney test). d Modulation of ATP-induced TO-PRO-3 uptake in HEK293T mP2RX7 cells (F1/F0) in cells treated with ATP and HEI3090 (25 nM). Error bars are mean ± SEM (n = 3 independent experiments, 4 replicates, two-tailed Mann–Whitney test). e Average fluorescence intensities in HEK293T mP2RX7 or control HEK pcDNA6 measured 10 min after stimulation with ATP and HEI3090 at concentrations of 25, 250, and 2.5 µM, as indicated in the color code. Data are presented as scatter dot plots ± SEM (n = 3 independent experiments and 4 replicates, two-tailed Mann–Whitney test). f Left: average Fluo-4-AM fluorescence intensities in WT or p2rx7−/− splenocytes stimulated with 50 µM ATP measured at the plateau i.e., 10 min after stimulation. Data are presented as scatter dot plots ± SEM (n = 2 independent experiments and 4 replicates). Right: graph represents the percentage of TO-PRO-3 positive cells in splenocytes isolated from naïve WT or p2rx7−/− mice. Data are presented as scatter dot plots ± SEM (n = 3 independent experiments in duplicate, two-tailed Man–Whitney test). g Pharmacokinetic analysis of HEI3090 intraperitoneally injected in WT mice. Error bars are means ± SEM (n = 3 independent experiments in duplicate). Bars are mean ± SEM. p values: *p < 0.05, **p < 0.01 ***p < 0.001, ****p < 0.0001. Source data are provided as a Source Data file.

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