Figure 5 Tacrolimus interacted directly with MRGPRX2 and regulated MRGPRX2.
(A) Molecular docking between tacrolimus and MRGPRX2.(B) Intracellular calcium concentrations of NC-LAD2 cells, MRGPRX2-knockdown LAD2 cells and HMC-1 cells treated by short-term tacrolimus treatment. Calcium images at different times of NC-LAD2 cells, MRGPRX2-knockdown LAD2 cells and HMC-1 cells were presented respectively, fluorescence intensity represents the intracellular Ca2+ concentration. (C) Western blot analysis of the expression levels of P-PLCγ1, PLCγ, P-PKC, PKC, P-ERK, ERK, P-P38, P38 and GAPDH in LAD2 cells, and quantification of P-PLCγ1, PLCγ, P-PKC, PKC, P-ERK, ERK, P-P38 and P38 protein expression respectively by densitometric analysis. Experiments of (B) and (C) were repeated 3 times. Statistical significance was accepted at p < 0.05 (*p < 0.05, **p < 0.01 and ***p<0.005) by two-tailed unpaired Student’s t-test.