Fig. 3

ACYP2 promotes glioma cell proliferation through altering intracellular Ca²⁺ homeostasis and calpain activity. a, Representative images showing free intracellular Ca²⁺ in the indicated cells under the confocal microscope (left panel). Green color represents staining of free intracellular Ca²⁺. Histogram represents mean ± SD of the fluorescence intensity from five microscopic fields in each group, as shown in right panel. Scale bars, 200 μm. b, Free intracellular Ca²⁺ measured by flow cytometer was shown in the middle and right panels. The data were presented as mean ± SD (n = 3). c, Cells were transfected with the indicated siRNAs and treated with the vehicle or 5 μM BAPTA-AM for 6 h, the MTT assay was then carried out to evaluate their effect on cell proliferation. The data were presented as mean ± SD (n = 3). d, Calpain activity was measured in the indicated cells using a calpain activity assay kit. Active Calpain I and calpain inhibitor Z-LLY-FMK were used as positive and negative control, respectively. Data are presented as mean ± SD (n = 3). e, Cells were transfected with the indicated siRNAs and treated with the vehicle or 10 μM calpeptin for 12 h, and the MTT assay was then carried out to evaluate their effect on cell proliferation. The data were presented as mean ± SD (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001