Fig. 5.

Manipulation of calcium voltage-gated channel subunit alpha1 C (Ca_V1.2) expression influences function of mast cell (MC) and HEK293 cell. (A) Ca_V1.2 expression was knockdown by small interfering RNA (siRNA). (B) β-hexosaminidase release induced by compound 48/80 (C48/80), substance P (SP), and ciprofloxacin (Cipro) in Ca_V1.2 knockdown laboratory of allergic diseases 2 (LAD2) cells. (C) Ca²⁺ influx induced by C48/80 (left) or SP (right) was in Ca_V1.2 knockdown cells. (D) Mas-related G protein-coupled receptor-X2 (MrgX2)⁺ cells and MrgX2⁺Ca_V1.2⁺ validation. (E) Ca²⁺ influx in HEK293 cells without MrgX2. (F) C48/80 induced Ca²⁺ influx in MrgX2⁺Ca_V1.2⁺ cells (left) and MrgX2⁺ cells (right). (G) Fluorescence of Ca²⁺ in C48/80 induced MrgX2⁺Ca_V1.2⁺ cells and MrgX2⁺ cells. (H) The effect of Nimodipine (Nim) on Ca²⁺ influx in MrgX2⁺Ca_V1.2⁺ cells (left) and MrgX2⁺ cells (right). Experiments were repeated three times. Data are presented as the mean ± standard error of mean (SEM) and were analyzed using two-tailed unpaired Student's t-test and one-way analysis of variance (ANOVA) test. ^∗P < 0.05 and ^∗∗P < 0.01, compared between experiment groups and vehicle or HEK293. NC: negative control; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; RFU: relative fluorescence unit.