TABLE 1.
Real-time PCR assays of VDR mRNA expression in Caco2, HEK293, HepG2, primary human hepatocytes and mouse livers
Primary human hepatocytes were isolated from seven human donors (indicated by HH#). Two micrograms of mRNA of Caco2, HEK293, HepG2, human hepatocytes, or mouse livers were used for the reverse-transcription reaction. UBC mRNA expression was monitored as the internal control. Ct values of VDR and UBC were measured by quantitative real-time PCR. Assays were done in triplicate. ΔCt was calculated by subtracting Ct of UBC from Ct of VDR. The S.D. of ΔCt = square root [(SD of Ct of UBC)2 + (SD of VDR)2].
|
mRNA Sample
|
VDR
|
|
|---|---|---|
| Ct Value ± S.D. | ΔCt Value ± S.D. | |
| Caco2 | 27.5 ± 0.13 | 7.8 ± 0.13 |
| HEK293 | 34.7 ± 0.17 | 13.2 ± 0.17 |
| HepG2 | 34.8 ± 0.05 | 13.9 ± 0.06 |
| HH1318 | 31.5 ± 0.14 | 10.9 ± 0.18 |
| HH1320 | 31.7 ± 0.13 | 12.3 ± 0.13 |
| HH1358 | 30.1 ± 0.07 | 10.8 ± 0.09 |
| HH1361 | 27.6 ± 0.06 | 7.8 ± 0.07 |
| HH1363 | 28.1 ± 0.11 | 9.0 ± 0.11 |
| HH1367 | 30.5 ± 0.06 | 8.4 ± 0.17 |
| HH1393 | 32.2 ± 0.01 | 11.9 ± 0.01 |
| Average human hepatocytes | 30.2 ± 1.79 | 10.2 ± 1.76 |
| Mouse 1 | 38.5 ± 0.08 | 19.0 ± 0.09 |
| Mouse 2 | 36.3 ± 0.1 | 18.1 ± 0.1 |
| Average mouse livers | 37.4 ± 1.56 | 18.6 ± 0.64 |