Figure 2
G protein-selective AT1 activation by RAGE ligands in transgenic CHO cells. (a) Dose-dependent response in inositol monophosphate (IP1) concentration in response to angiotensin II (10–12–10–5 M) in CHO-RAGE-AT1. The measurement is expressed as % of IP1 concentration in the vehicle treatment. (number(n) = 4, each). (b) IP1 concentration in response to vehicle, 0.3 µg/mL HMGB1, 100 µg/mL AGE-BSA, or 10–10M angiotensin II in genetically engineered CHO cells. The concentration in the control wells of each cell type was normalized to 100% (n = 8 for vehicle, HMGB-1, and AGE-BSA, and n = 4 for angiotensin II in CHO-RAGE-AT1, n = 4 in CHO-RAGE-AT1mt) *p < 0.01 vs. vehicle treatment. (c) Dose-dependent response in cyclic adenosine monophosphate (cAMP) concentration in response to HMGB1(10–4–101 µg/ml) or angiotensin II (10–12-10–6 M) in CHO-RAGE-AT1. Cells were treated with 1 µM forskolin to induce cAMP accumulation. n = 4, each). (d) cAMP concentration in response to vehicle ,0.3 µg/mL HMGB1, 100 µg/mL AGE-BSA, or 10–10 M angiotensin II in genetically engineered CHO cells. Gi inhibitor, pertussis toxin (PTX), was pre-treated at 25 ng/mL for 12 h before stimulation. The concentration in control-treated wells in each cell type was normalized to 100% (n = 10 in CHO-RAGE-AT1, and n = 7 in CHO-RAGE, CHO-RAGE-AT1 with PTX treatment, CHO-RAGE-At1mt, RAGE-AT2, and RAGE-mDia-1) *p < 0.01 vs. vehicle treatment. (e) cAMP concentration in response to vehicle, 0.3 µg/mL HMGB1, 100 µg/mL AGE-BSA, or 10–10 M angiotensin II in CHO-RAGE-AT1 pre-treated with vehicle, 10 µM losartan (AT1 blocker), or 10 µM FPS-ZM1 (RAGE inhibitor) (n = 8, each). *p < 0.01 vs. vehicle treatment. The differences were determined by one-way ANOVA with Bonferroni correction.
