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Fig. 5 | Particle and Fibre Toxicology

Fig. 5

From: Nitrogen-doped graphene quantum dots induce ferroptosis through disrupting calcium homeostasis in microglia

Fig. 5

N-GQDs increased intracllular calcium level through activating calcium channels of L-VGCC and RyR in BV2 cells. a KEGG signaling pathways enriched by differentially expressed genes caused by 100 µg/mL N-GQDs in BV2 cells for 24 h using microarray analysis; b the levels of calcium in hippocampus; c, d representative fluorescent images of intracellular calcium level in BV2 cells showed green using Fluo4 dye with HBSS containing calcium and magnesium or HBSS containing EDTA without calcium and magnesium. The nucleus showed blue using DAPI, Scale bar: 20 µm; e heatmap of differentially expressed genes associated with calcium signaling pathway in BV2 cells treated with 100 µg/mL N-GQDs for 24 h using microarray analysis; f the protein expressions of CACNA1D and RYR2 in BV2 cells using western blotting analysis; g, h representative fluorescent images of intracellular calcium level in BV2 cells showed green using Fluo4 dye with HBSS containing calcium and magnesium or HBSS containing EDTA without calcium and magnesium. The nucleus showed blue using DAPI, Scale bar: 20 µm. Each mouse was intranasally instilled with saline, 0.1 and 1 mg/kg BW N-GQDs every other day for 28 days (mouse n = 6). BV2 cells were treated with 25, 50 and 100 μg/mL N-GQDs for 24 h or BV2 cells were pre-treated with nifedipine, diltiazem, verapamil and dantrolene for 2 h and then exposed to 100 µg/mL N-GQDs for 24 h (n = 3). Data are showed as mean + SD of three independent experiments. Data are showed as mean ± SD of three independent experiments. The one-way ANOVA followed by the Dunnett’s t test were used to determine statistical significance (*P < 0.05, **P < 0.01, ***P < 0.001 vs. the control)

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