Journal: Human Molecular Genetics

Article Title: The biological impact of blood pressure-associated genetic variants in the natriuretic peptide receptor C gene on human vascular smooth muscle

doi: 10.1093/hmg/ddx375

Figure Lengend Snippet: The BP-elevating variant at NPR3 locus is associated with increased human VSMCs proliferation. VSMCs in the absence and presence of C-type natriuretic peptide (CNP; 100 nmol/l) and the NPR-C specific agonist (cANF 4–23 ; 100 nmol/l) for 24 h were analysed for proliferation rate. Cell proliferation over the 24-h period was calculated as (OD450 at 24 h−OD450 at baseline)/OD450 at baseline. ‘E’ indicates the BP-elevating allele, ‘L’ denotes the BP-lowering allele. ( A ) Differences in proliferation between untreated cells of different genotypes. Representative images of DAPI stained cells of the three different genotypes at baseline and 24 h. Scale bars=200µm. Mean±SEM, * P <0.05. ( B ) Differences in proliferation between CNP or cANF 4–23 treated cells of different genotypes, and ( C ) Inhibitory effects of CNP and cANF 4–23 on proliferation of cells of different genotypes, calculated by (OD450 at 24 h of untreated cells−OD450 at 24 h of CNP-treated or cANF 4–23 -treated cells)/OD450 at 24 h of untreated cells. Mean±SEM, * P <0.05.

Article Snippet: After overnight incubation, VSMCs were serum starved for 24 h then transferred to 15% FBS DMEM with or without 100 nmol/l CNP (Sigma-Aldrich) or cANF 4–23 (Bachem).

Techniques: Variant Assay, Staining

Journal: Human Molecular Genetics

Article Title: The biological impact of blood pressure-associated genetic variants in the natriuretic peptide receptor C gene on human vascular smooth muscle

doi: 10.1093/hmg/ddx375

Figure Lengend Snippet: The BP-elevating allele at NPR3 locus increases Ang II-induced intracellular calcium changes in human VSMCs. Human VSMCs were incubated with or without C-type natriuretic peptide (CNP; 1 μmol/l) and the NPR-C specific agonist cANF 4–23 (1 μmol/l), and then exposed to Ang II (10 or 100 nmol/l). ‘E’ indicates the BP-elevating allele, ‘L’ denotes the BP-lowering allele. ( A ) Representative images of intracellular calcium changes among genotypes under stimulation of 100 nmol/l Ang II. ( B ) Differences between genotypes in intracellular calcium changes in response to Ang II in the absence of CNP or cANF 4–23 . Differences between genotypes in intracellular calcium level changes in response to Ang II ( C ) and drug inhibitory effect ( D ) in the presence of CNP or cANF 4–23 . Mean±SEM, * P <0.05, ** P <0.01.

Article Snippet: After overnight incubation, VSMCs were serum starved for 24 h then transferred to 15% FBS DMEM with or without 100 nmol/l CNP (Sigma-Aldrich) or cANF 4–23 (Bachem).

Techniques: Incubation

Journal: Diabetologia

Article Title: GPR120 (FFAR4) is preferentially expressed in pancreatic delta cells and regulates somatostatin secretion from murine islets of Langerhans

doi: 10.1007/s00125-014-3213-0

Figure Lengend Snippet: Effects of selective GPR120 agonists on somatostatin secretion from isolated mouse islets of Langerhans. Mouse islets were isolated, cultured overnight and incubated for 2 h with 3 mmol/l or 16.6 mmol/l glucose in the absence or presence of 100 nmol/l Glip, 500 μmol/l CCh or 30 μmol/l of each of three GPR120 agonists (Metabolex 36 [Metab 36], AZ-423 and AZ-670). The incubation medium was supplemented with 500 μmol/l IBMX. Following incubation, the supernatant fraction was collected and analysed for somatostatin secretion by ELISA. Data are presented as mean values ± SEM and the experiment was repeated on a minimum of three separate occasions. ** p < 0.01 vs 3 mmol/l glucose alone; *** p < 0.001 vs 3 mmol/l glucose alone; †† p < 0.01 vs 16.6 mmol/l glucose alone

Article Snippet: Mouse islets were isolated, cultured overnight and incubated for 2 h with 3 mmol/l or 16.6 mmol/l glucose in the absence or presence of 100 nmol/l Glip, 500 μmol/l CCh or 30 μmol/l of each of three GPR120 agonists (Metabolex 36 [Metab 36], AZ-423 and AZ-670).

Techniques: Isolation, Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay