Fig. 3.
Calcium voltage-gated channel subunit alpha1 C (CaV1.2) was activated by phosphorylating at Ser1928 through protein kinase C (PKC). (A) The membrane potential stimulated by different Mas-related G protein-coupled receptor X2 (MrgX2) agonists. (B) p-CaV1.2 at Ser1928 when induced by compound 48/80 (C48/80) and treated with nimodipine (Nim) in laboratory of allergic diseases 2 (LAD2) cells. (C) The phosphorylation of PKC and protein kinase A (PKA) induced by C48/80 and treated with Ro 31-8220 (Ro) (left) or PKA inhibitor (PKI) (right). (D) Ca2+ influx (left) and degranulation (right) in LAD2 cells treated with Ro or PKI. (E) Pro-inflammatory cytokines release in LAD2 cells treated with Ro or PKI. (F) Co-immunoprecipitation (Co-IP) displayed between p-PKC and CaV1.2 in C48/80 induced LAD2 cells (left) and the effect of Ro on the direct interaction (right). Experiments were repeated three times. Data are presented as the mean ± standard error of mean (SEM) and were analyzed using one-way analysis of variance (ANOVA) test. ∗∗∗P < 0.001, compared between experiment and vehicle or negative control (NC) groups; ###P < 0.001, compared between experiment and NC groups. SP: substance P; Sino: sinomenine; RFU: relative fluorescence unit; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; TNF-α: tumor necrosis factor; CCL-2: C–C motif chemokine ligand 2.