PKIα expression does not affect AT1-R mRNA or protein levels in VSMCs
| Antet | Wild-Type | eGFP | PKIα-eGFP | |||
|---|---|---|---|---|---|---|
| (+) | (−) | (+) | (−) | (+) | (−) | |
| AT1-R mRNA levels | 100 ± 9 | 100 | 103 ± 24 | 66 ± 15 | 108 ± 22 | 66 ± 14 |
| Mean ± standard error (n) | (2) | (3) | (3) | (3) | (3) | (3) |
| AT1-Receptor levels | ||||||
| B max (fmol/mg of protein) | 177 | 181 | 161 ± 32 | 141 ± 42 | 120 ± 18 | 150 ± 24 |
| K D(nm) | 0.27 | 0.30 | 0.30 ± 0.05 | 0.30 ± 0.06 | 0.35 ± 0.09 | 0.28 ± 0.08 |
| Mean ± standard error (n) | (3) | (3) | (3) | (3) | ||
Wild-type VSMCs and cells expressing either GFP or PKIα-eGFP were cultured with (+) or without (−) 0.1 μg/ml Antet for 24 hr before cell membranes were prepared and subjected to saturation binding with125I-sarile as described in Experimental Procedures.B max and K D were determined by 5-point nonlinear regression using one-site binding equation from GraphPAD Prizm software. AT1-R mRNA levels were determined by Northern hybridization of 10 μg of total RNA separated on a formaldehyde gel with radiolabeled AT1-R cDNA probe. AT1-R mRNA levels are normalized as percentage of that in wild-type VSMCs without Antet. One-way analysis of variance indicates there are no significant differences (p > 0.05) in the measured parameters across the various cell lines.