Intracellular FVIII:Ag and FVIII mRNA
| . | HCMECs . | HUVECs . |
|---|---|---|
| FVIII:Ag prepared by lysis buffer, mU/well* | 2.3 ± 0.3 (n = 5) | ND |
| FVIII:Ag prepared by F/T buffer, mU/well* | 1.3 ± 0.1 (n = 5) | ND |
| VWF:Ag prepared by lysis buffer, mU/well* | 20.1 ± 2 (n = 5) | 31.6 ± 3.4 (n = 10) |
| VWF:Ag prepared by F/T buffer, mU/well* | 7.3 ± 0.6 (n = 5) | ND |
| FVIII:C prepared by lysis buffer, mU/well* | 2.5 ± 0.4 (n = 5) | < 0.2 (n = 10) |
| FVIII:C prepared by F/T buffer, mU/well* | 0.9 ± 0.1 (n = 5) | < 0.2 (n = 10) |
| FVIII mRNA† | 94.3 ± 0.1 | 0 |
| VWF mRNA† | 394 714.5 ± 0.1 | 489 161 ± 0.2 |
| . | HCMECs . | HUVECs . |
|---|---|---|
| FVIII:Ag prepared by lysis buffer, mU/well* | 2.3 ± 0.3 (n = 5) | ND |
| FVIII:Ag prepared by F/T buffer, mU/well* | 1.3 ± 0.1 (n = 5) | ND |
| VWF:Ag prepared by lysis buffer, mU/well* | 20.1 ± 2 (n = 5) | 31.6 ± 3.4 (n = 10) |
| VWF:Ag prepared by F/T buffer, mU/well* | 7.3 ± 0.6 (n = 5) | ND |
| FVIII:C prepared by lysis buffer, mU/well* | 2.5 ± 0.4 (n = 5) | < 0.2 (n = 10) |
| FVIII:C prepared by F/T buffer, mU/well* | 0.9 ± 0.1 (n = 5) | < 0.2 (n = 10) |
| FVIII mRNA† | 94.3 ± 0.1 | 0 |
| VWF mRNA† | 394 714.5 ± 0.1 | 489 161 ± 0.2 |
ND indicates not done.
Cell extracts of 5 microculture wells were prepared either by lysis buffer or by freeze/thawing (F/T) buffer. FVIII:Ag and VWF:Ag were measured with ELISA; FVIII:C was measured with a FVIII chromogenic assay. Results are expressed as the mean plus or minus SD of 5 or 10 microculture wells.
FVIII and VWF mRNA levels in HCMECs and HUVECs are normalized to the level of corresponding genes in a control cell line with the use of the 2−ΔΔCt method. GAPDH expression served as the control gene. All assays were performed in duplicate.