Fig. 4: Reduced sphere growth at pH 6.6 is ASIC1a-dependent.
From: Acidosis induces RIPK1-dependent death of glioblastoma stem cells via acid-sensing ion channel 1a
a–f SFR after 12 days at pH 7.4 or at pH 6.6 with 100 nM PcTx1, 500 nM APETx2, or 100 nM PcTx1 and 20 µM Nec-1 combined. SFR was normalized to DMSO pH 7.4 control, which was 74.5 ± 11.1 for a, 46.3 ± 6.7 for b, 102.4 ± 11.5 for c and e, and 77 ± 7.9 for d and f. 1 data point represents 1 well. n ≥ 17 wells per condition, 2 independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. One-way ANOVA was followed by Dunnett’s post-hoc test. g Traces of whole-cell patch clamp experiments with wildtype or ASIC1a knockout R54 cells. ASIC currents were elicited by a pH drop from pH 7.3 to 6.0. Each trace is representative of five similar measurements. h SFR after 12 days at pH 7.4 or at pH 6.6. SFR was normalized to the respective pH 7.4 control, which was 104.7 ± 11 for wt R54, 77 ± 9.1 for KO1, and 60.4 ± 6.8 for KO2. n = 24 wells per condition from 2 independent experiments. i Sphere diameters (mean ± SD) in µm of 100 spheres per condition after 12 days at pH 7.4 or at pH 6.6. Measurements are from 2 independent experiments. Data points represent single sphere diameters. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 (one-way ANOVA followed by Sidak’s post-hoc test).
