Table 1.
Characteristics of biological and technical replicates* utilized in the microarray analysis
| Sample ID |
Cell/Tissue Type | Experimental Condition |
Measure of AT1R Activation/Inhibition |
|---|---|---|---|
| SK-1 | HEK-AT1R | Untreated | Control |
| SK-2 | HEK-AT1R | AngII | ERK1/2, JAK, STAT3 phosphorylation |
| SK-3 | HEK-AT2R | Untreated | Control |
| SK-4 | HEK-AT2R | AngII | – |
| SK-5 | HASMC | Untreated | Control |
| SK-6 | HASMC | AngII | ERK1/2 phosphorylation |
| SK-7 | HASMC | Losartan | ERK1/2 phosphorylation |
| SK-8 | HASMC | Untreated | Control |
| SK-9 | HASMC | AngII | ERK1/2 phosphorylation |
| SK-10 | HASMC | Losartan | ERK1/2 phosphorylation |
| SK-11 | HL1-AT1R | U ntreated | Control |
| SK-12 | HL1-AT1R | AngII | ERK1/2, JAK, STAT3 phosphorylation |
| SK-13 | RASMC | Untreated | Control |
| SK-14 | RASMC | AngII | ERK1/2 phosphorylation |
| SK-15 | RASMC | Losartan | ERK1/2 phosphorylation |
| | SK-16 | RASMC-AT1R | Untreated | Control |
| SK-17 | RASMC-AT1R | AngII | ERK1/2, STAT3 phosphorylation |
| SK-18 | RASMC-AT2R | Untreated | Control |
| SK-19 | RASMC-AT2R | AngII | – |
| SK-21 | Whole heart (C3H NT) |
Non- transgenic |
Control |
| SK-22 | Whole heart (C3H TG) |
Cardiac- specific AT1R transgene overexpression |
ERK1/2, JAK, STAT3 phosphorylation, Cardiac Hypertrophy & HF Phenotype |
| SK-23 | Whole heart (C57BL/6 NT) |
Non- transgenic |
Control |
| SK-24 | Whole heart (C57BL/6 TG) |
Cardiac- specific AT1R transgene overexpression |
ERK1/2, JAK, STAT3 phosphorylation, Cardiac Hypertrophy & HF Phenotype |
Details regarding each RNA sample utilized for miRNA expression profiling are shown. For primary comparison, correlations were made between the untreated and ligand treated cells or between NT and TG heart tissue samples. The samples include HEK-293 cells expressing AT1R or AT2R, an immortalized rat aortic smooth muscle cell (RASMC) line expressing AT1R or AT2R and a mouse atrial cell line (HL-1) expressing AT1R. Two independent human aortic smooth muscle cell (HASMC) samples were profiled to reduce potential bias that may result from the variability in primary cells (passage # 2-3). The heart tissue analyzed was from AT1R overexpressing TG mice in which cardiac hypertrophy and HF phenotype were assessed by 1) measuring the heart weight to body weight ratio, 2) measuring ejection fraction and wall thickness by echocardiography (to monitor ejection fraction and wall thickness), and 3) expression of BNP as an independent marker of heart failure as reported previously [34]. *Biological replicates control for biological diversity of a response, measure a quantity from different sources under the same condition and are often more powerful. Biological replicates in this study are defined as RNA isolated independently from multiple sources, each with a distinct genome under the same condition, AngII-activation of the AT1R. Technical replicates are defined as RNA isolated from an AngII-responsive source with same genome under different treatment conditions.