Fig. 2: Calcium and CaSR impact leukaemia cell function.
From: Distinct and targetable role of calcium-sensing receptor in leukaemia
a Percentage of CaSR+ cells of all cells in different human leukaemia cell lines (one-way ANOVA, Dunnett test, mean ± SD) (K562: BCR-ABL1+ (CML, n = 9), NALM-6: B-ALL (n = 3), THP1: MLL-AF9 (AML, n = 8), Kasumi: AML1-ETO (AML, n = 3)). n refers to biological replicates. b, c Representative immunoblot (b) and its quantification (c) (mean ± SD) for CaSR and GAPDH expression in lysates of THP1 cells treated with 0–7.5 mM of CaCl2 for 4 h. The blot is representative of three independent experiments. d, e Representative immunoblot (of three independent experiments) (d) and its quantification (e) (mean ± SD) for CaSR and GAPDH expression in K562 cells treated with CaCl2. f, g Calcium flux analysis of THP1 (f) and K562 (g). h, i Number of THP1 (h) or K562 (i) cells adhering to fibronectin (FN). Prior to adhesion, cells had been treated with calcium (n = 6, one-way ANOVA, Dunnett test, mean ± SD). j, k Number of THP1 (j) or K562 (k) cells which migrated towards C-X-C motif chemokine 12 (CXCL12). Cells had been pre-treated with calcium (n = 4, one-way ANOVA, Dunnett test, mean ± SD). From h–k, n refers to biological replicates. l–o Distance (in µm) of intravenously transplanted CMTMR-labelled THP1 (l, m) or K562 (n, o) cells to the endosteum in the calvarium of NOD SCID interleukin-2 receptor γ (IL2Rγ)−/− (NSG) mice. NTC (non-target control) (black) versus CaSR knockout (KO) (blue) cells (l, n) and THP1 WT (black) versus CaSR-overexpressing (OE) THP1 or K562 (red) (m,o) cells had been transplanted and imaged by intravital microscopy (two-tailed t test, mean ± SD) (n = 30 in (l), n = 29 (THP1 WT), n = 30 (THP1 CaSR OE) in m, n = 27 (K562 NTC), n = 53 (K562 CaSR KO) in n, n = 40 (K562 WT), n = 50 (K562 CaSR OE) in o. p, q Distance of transplanted THP1 (p) or K562 (q) cells to the endosteum. Cells had been pre-treated with NPS-2143. Experiments were performed in three different mice per cohort (n = 35 (vehicle), n = 38 (NPS-2143) in p, n = 30 (vehicle), n = 42 (NPS-2143) in q). From l–q n refers to individual cells. Data are presented as mean values ± SD. Source data are provided as a Source Data file.
