Fig. 5: CBD induces cytosolic Ca2+ rise in human T-ALL cell lines and non-cancerous T cells.
a–c [Ca2+]i monitoring in leukemic cells, treated with CBD. Traces represent the mean ± SD of at least six samples from independent experiments. d Dose dependence of peak [Ca2+]i values in T-ALL cell lines, exposed to CBD. Data are mean ± SD for at least six samples from independent experiments. e, f [Ca2+]i monitoring in non-cancerous T cells in resting (e) and activated (f) states. Traces represent the mean ± SD of at least six samples from independent experiments. g Pharmacological analysis of [Ca2+]i rise in response to CBD. Before CBD treatment, cells were preincubated over 20 min with either CB1 antagonist, rimonabant (1 μM), CB2 inverse agonist, AM630 (2 μM), or GPR55 antagonist, CID16020046 (3 μM); values Δ [Ca2+]i were obtained by subtracting the [Ca2+]i baseline level from [Ca2+]i maximum increase. Data are mean ± SD of at least 6 samples from independent experiments. h [Ca2+]i monitoring in Jurkat cells, suspended in Ca2+-free HBSS. Traces represent the mean ± SD of at least six samples from independent experiments. i, j Effect of Gd3+ and RR, non-selective blockers of plasma membrane Ca2+- permeable channels, on [Ca2+]i (i and j, left) and cell viability evaluated by resazurin-based metabolic assay (i and j, right). Gd3+ (1 μM) or RR (1 μM) were added to Jurkat cells 20 min before the [Ca2+]i measurement. i, j left: traces represent the mean ± SD of at least six samples from independent experiments. i, j right: data (resorufin fluorescence intensity) were normalized to vehicle-treated control and reported as mean ± SD (n = 8 of four independent experiments (****p < 0.0001, Student’s t-test)
