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. 2016 Dec 16;11(12):e0167622. doi: 10.1371/journal.pone.0167622

Fig 5. Analysis of TRPV1-dependent calcium trafficking in human spermatozoa.

Fig 5

A)Western blot analysis of TRPV1 expression in isolated sperm cytoplasm and sperm membrane obtained by differential centrifugation. β-Actin was used as cytoplasm marker. Images are representative of three independent experiments. D) Dynamic of intracellular calcium concentration in sperm cells stimulated with 10 μM capsaicin (CPS), CPS pre incubated with 10 μM capsazepine (CPS + CPZ), CPS pre incubated with 2,5 μM rabbit polyclonal anti-TRPV1 antibody (CPS + Ab), CPS with chelation of extracellular calcium obtained with addition of 6mM EDTA (CPS + EDTA) compared to unstimulated conditions (CTRL). In C) values of area under the curves normalized on controls (Normalized A.U.C.) are compared. Significance: * = P<0,05 vs CTRL B) Dynamic of intracellular calcium concentration in sperm cells stimulated with 10 μg/mL progesterone (P4), P4 pre incubated with 10 μM capsazepine (P4 + CPZ), P4 pre incubated with 2,5 μM rabbit polyclonal anti-TRPV1 antibody (P4 + Ab), P4 with chelation of extracellular calcium obtained with addition of 6mM EDTA (P4 + EDTA) compared to unstimulated conditions (CTRL). In E) values of area under the curves normalized on controls (Normalized A.U.C.) are compared. Significance: * = P<0,05 vs CTRL