Extended Data Fig. 7: Vortioxetine induces a robust calcium response and alters the electrophysiological properties of glioblastoma cells.
a, ER calcium store release measured by FLIPR assays in LN-229 cells (n = 4 assay plates; n = 18 conditions; n = 12 wells/drug; DMSO and Thapsigargin (TG) positive control, n = 24 wells each). b, Extracellular calcium influx measured by FLIPR assays in F050.C (n = 17 conditions as in Fig. 5f). (*) denote conditions where the median [Ca2+ fold change] < 0. Black line: median value. a, b, Fold change relative to DMSO after drug treatment. Two-tailed t-test against DMSO. P-values adjusted for multiple comparisons by Holm correction. Black line: median value. c, Single-cell-resolved calcium response (ΔF/F0) measured by ratiometric Fura-2 imaging across 6 cell lines (x-axis; n = 3,561 cells total). Mean change in calcium signal immediately after Vortioxtine treatment compared to baseline, each averaged across a 120 s time window. Paired (baseline vs drug treatment) two-tailed Wilcoxon test. d, Calcium response type stratified by the presence (Ψ) or absence (Ø) of oscillatory calcium signaling at baseline (BASE) and Vortioxetine (VORT; 20) treatment. ‘VORT 1-2 Peaks’: non-oscillatory calcium response with 1-2 peaks after VORT treatment. e, Heatmap of single-cell-resolved (rows) calcium response (ΔF/F0) for response type ‘VORT 1-2 Peaks’ across time (x-axis). NR: no response. f, Max (top) and mean (bottom) peak amplitude of ΔF/F0 for type ‘BASE Ψ, VORT Ψ’ (n = 501 cells) displaying oscillatory calcium signaling during both time spans across the 6 cell lines in d. Paired two-tailed t-test. g, Resting membrane potential (Vm) of LN-229 (n = 13) and LN-308 cells (n = 10) measured by whole-cell patch-clamp before (CTRL) and after VORT treatment (10 µM) in matched single-cells (connected by grey lines). Paired two-tailed t-test. h, Representative single-cell current traces for each cell line (LN-229, LN-308) and condition (CTRL, VORT) corresponding to the voltage-clamp protocol (legend). i, Current-voltage characteristics (I-V curves) of LN-229 (n = 13) and LN-308 cells (n = 10) in g, before (CTRL) and after VORT treatment (10 µM) in matched single-cells. Standard error of the mean (SEM) shown as error bars. See Methods for description of summary statistics. j, Relative gene (panels) expression upon siRNA knockdown (columns) normalized to the FLUC negative control siRNA (n = 3 biological replicates; dots). Two-tailed t-test with adjusted P-values after Holm correction. Boxplots as in Fig. 1b.
