Skip to main content
. 2018 Oct 23;4:57. doi: 10.1038/s41421-018-0058-6

Fig. 3. Functional analysis of the ADRB2 Arg16Gly variants in cell culture systems.

Fig. 3

a The western blots of β2AR and GAPDH for lysates of adenovirus-infected rat cardiomyocytes showing that the transfection method (multiplicity of infection of 100 for 24 h) produced equal expression of ADRB2-Arg16 and ADRB2-Gly16, which is true irrespective of deglycosylation treatment of the cell lysates with PNGase F. b PTX treatment (0.75 μg/ml overnight) enhanced the zinterol (1×10−5 M)-induced contractile response in adult rat cardiac myocytes expressing ADRB2-Arg16, but not in those expressing ADRB2-Gly16 (multiplicity of infection of 100 for 24 h; N = 8 for each group). c ADRB2 knockout mouse cardiomyocytes were infected with Adeno-ADRB2-Arg16 or Adeno-ADRB2-Gly16 viruses as described in Materials and Methods and then subjected to stimulation with zinterol (3×10−7 M). The zinterol-induced contractile responses were 200−300% in the absence of PTX. PTX enhanced these responses in cells expressing ADRB2-Arg16, but not in cells expressing ADRB2-Gly16. (N = 10–16 cells from four mouse hearts for each data point). d Peripheral lymphocytes were isolated from HF patients with different genotypes at the ADRB2 Arg16Gly locus. The inhibition of Gi by PTX enhanced β2AR-mediated cAMP accumulation in cells with the AA genotype (N = 10; P < 0.05), whereas cAMP accumulation in cells with either the AG (N = 10) or GG (N = 6) genotype was insensitive to PTX treatment. Therefore, peripheral lymphocytes harboring the G allele of the ADRB2 Arg16Gly A > G polymorphism showed deficient coupling of Gi to the β2AR. All the data were shown as the mean ± SEM. β2AR β2 adrenergic receptor, HF heart failure, PTX pertussis toxin. *p<0.05, **p<0.01, ***p<0.001