Fig. 3: TRPA1 mediates H2O2-induced intracellular Ca2+ signals in primary cultures of mCRC cells. | Cell Death Discovery

Fig. 3: TRPA1 mediates H2O2-induced intracellular Ca2+ signals in primary cultures of mCRC cells.

From: Transient receptor potential ankyrin 1 (TRPA1) mediates reactive oxygen species-induced Ca2+ entry, mitochondrial dysfunction, and caspase-3/7 activation in primary cultures of metastatic colorectal carcinoma cells

Fig. 3: TRPA1 mediates H2O2-induced intracellular Ca2+ signals in primary cultures of mCRC cells.

A H2O2 (50 µM) induces larger Ca2+ signals in mCRC as compared to non-neoplastic cells. B Mean ± SE of peak Ca2+ signal evoked by H2O2 in both mCRC and non-neoplastic cells. Student’s t-test: ***p < 0.001. The placed above the scattered dots represent the number of responding cells out of the total cell number. N = 4 for each experimental condition. C Intracellular Ca2+ signals induced by 50 µM H2O2 (Ctrl) were abrogated in mCRC cells pretreated with the specific TRPA1 inhibitor, HC-030031 (30 µM, 30 min), or transfected with the selective siTRPA1. D Mean ± SE of the peak Ca2+ signal evoked by H2O2 in control (Ctrl) mCRC cells and in mCRC cells transfected with siTRPA1 or pretreated with HC-030031. One-way ANOVA followed by the post hoc Dunnett’s test: ***p < 0.001. The numbers placed above the scattered dots represent the number of responding cells out of the total cell number. N = 4 for each experimental condition. E Intracellular Ca2+ signals induced by 50 µM H2O2 in the absence (Ctrl) and presence of the thiol-reducing compound, DTT (5 µM), the H2O2 scavenger, catalase (500 U/mL) or the iron-chelating compound, deferoxamine (100 µM). F Mean ± SE of peak Ca2+ signal evoked by H2O2 under the designated treatments. The numbers placed above the scattered dots represent the number of responding cells out of the total cell number. N = 4 for each experimental condition. G Intracellular Ca2+ signals induced by the selective TRPA1 agonist, 4-HNE (30 µM), in mCRC cells under control conditions (Ctrl) or upon pharmacological (HC-030031; 30 µM, 30 min) or genetic (with a selective siTRPA1) blockade of TRPA1 activity. H Mean ± SE of peak Ca2+ signal evoked by 4-HNE in control (Ctrl) mCRC cells and in mCRC cells treated with DTT, catalase, and deferoxamine. One-way ANOVA followed by the post hoc Dunnett’s test: ***p < 0.001. The numbers placed above the scattered dots represent the number of responding cells out of the total cell number. N = 4 for each experimental condition.

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