This medium was supplemented with 4 ng/ml bFGF (GF003-AF, Millipore) and conditioned on irradiated mouse embryonic fibroblasts (MEFs) overnight, then filtered in a 0.1 µm PES filter (Fisher) before being supplemented with an additional10 ng/ml bFGF.
vCPH was concentrated to 30 mg/mL in 100 mM HEPES (pH 7.5) before being supplemented with 1.1 mM MnCl2.
Cells were incubated for 4 h at 37°C/5% CO2 before being supplemented with FBS to a final concentration of 10%.
Initial samples for determination of colony-forming ability of S. aureus present in culture medium and internalized by macrophages, as well as samples for microscopic examinations, PCR analysis, and hMDMs viability assessment were all taken immediately after cells were washed out of free non-phagocytosed bacteria, before medium was supplemented with gentamycin.
After boiling, food was allowed to cool down to 60°C 65°C before it was supplemented with 0.5% Bromophenol blue sodium salt (B5525, Sigma), 0.5% Bromocresol purple sodium salt (17492, Sigma), and/or 0.03% Fluoropoo (4,4-difluoro-1,3,5,7,8-pentamethyl-4-bora-3a,4a-diaza-s-indacene, BODIPY 493/503, Invitrogen).
A few other successes are ascribed to such observation before the balloon was supplemented by the far more valuable airplane in World War I. Nevertheless, the balloon never fulfilled its potential as a warning device.
The cells were grown for 48 60 h in the presence of viral particles before the medium was supplemented with 0.35 0.4 μg/ml puromycin (Sigma) for transgene selection.
Before final centrifugation, reactions were supplemented to 0.5 M NaCl for 5 min, then separated to yield pellets (P, vi and viii) and supernatants (S, v and vii).
In experiment 1, aliquots from ejaculates were supplemented before freezing with one of four levels of hyaluronan: 100 μg/mL, 200 μg/mL, 400 μg/mL, and 1000 μg/mL along with an untreated control.
Whenever DNA and DNase I effects were tested, the medium was supplemented before biofilm seeding (t = 0).
Zebrafish were fed twice daily with Zeigler Adult Zebrafish Diet (Aquatic Habitats) and on afternoons before breeding the diet was supplemented with live Artemia.
