Table 6.
Amount of TGFβ ligands present in rat serum and VIC media and effect of 48-h TGFβ neutralizing antibody or TGFβRi treatment on secreted ligands into VIC media
| TGFβ1 | TGFβ2 | TGFβ3 | |
|---|---|---|---|
| Control (pg/mL) | |||
| Rat serum | 5302.8 ± 191 | 3336.7 ± 93 | 187.5 ± 11.3 |
| Media alone (0.5% FBS/M199)a | 624.5 ± 20.3 | 22.21 ± 2.3 | <BDL |
| Conditioned mediab | 3558 ± 61.3 | 1676.6 ± 56.8 | <BDL |
| Neutralization antibodies (percentage of controld) | |||
| PBS control | 100 | 100 | <BDL |
| 1D11 (10 μg/mL) | ≤0 | 45 | NA |
| TGFβ1 (10 μg/mL) | 57.4 | 72.5 | NA |
| TGFβ2 (10 μg/mL) | 77.4 | 26.4 | NA |
| TGFβ3 (10 μg/m)L | 65.5 | 51.5 | NA |
| TGFβ2+TGFβ3 (10 μg/mL each) | 100.1 | 29.9 | NA |
| TGFβRis (percentage of controld) | |||
| DMSOc | 100 | 100 | <BDL |
| LY2109761 (10 μM) | 55 | 27.5 | NA |
| BMT-B (10 μM) | 59 | 31.9 | NA |
<BDL, below detectible level; NA, not applicable due to lack of detectable ligand in cell media.
Average of three experiments ± SEM.
Baseline TGFβ concentration was measured with pre-test culture media containing 0.5% FBS in M199.
PBS in 0.5% FBS/M199 was used as an experimental control for VICs treated with TGFβ neutralizing antibodies. The amount of TGFβ in these samples reflects 4–5 days' accumulation (seeding + starving).
0.05% DMSO in 0.5% FBS/M199 is used as an experimental control for VICs treated with TGFβRis.
Percent change calculation: (actual concentration – media alone/control concentration (PBS or DMSO)) × 100.