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Adherent cells were cultured in 96-well plates and 48 hours after transfection cells were first stimulated with AVP in DMEM-serum free supplemented by BSA 0.1% and then washed with twice PBS.

Cells were seeded in serum free, supplemented RPMI 1640 containing 0.2% fatty acid free BSA at 5 × 105 cells per ml/well in 24 well plates.

Cells were incubated with phenol red free supplemented DMEM at 37°C for 10 min with anti-CD81 Fab-Atto647N fragments (3 pM).

HELN-Fα/β cells were routinely grown in DMEM-F12 (phenol red free) supplemented with 3% of dextran-coated charcoal-treated fetal calf serum (DCC medium) and serum was deprived 24 hr before kinetics ChIP analysis.

GFP.Vpr-labelled HCVpp were incubated with phenol red free supplemented DMEM at 37°C for 10 min with anti-HCV E2 11/20-Atto647N Fab fragments (5 pM) prior to cell inoculation.

Six hr after the transfection, the culture medium was replaced with RPMI (phenol red free) supplemented with 10nM androstenedione and 5% charcoal-dextran treated fetal bovine serum (Biotechnics Research, CA USA).

Following shock, the medium was replaced with Phenol Red-free, supplemented DMEM containing 100 µM D-Luciferin, sealed with a custom-made glass coverslip (#1 borosilicate glass, 0.12 0.16 mm thickness, 115×77 mm area, Erie Scientific, Portsmouth, NH) and subjected to real-time bioluminescence measurements for 5 days.

This stock was then added to serum-free supplemented DMEM and frozen down in aliquots at 10 μg.mL-1.

Briefly, cortices were dissected from 15- to 16-day-old C57BL/6 mouse embryos and mechanically dissociated in PBS (Ca2+- and Mg2+-free) supplemented with glucose (33 m m).

We cultured MCF-7 and MDA-MB-231 cell lines in RPMI-1640 medium (phenol red-free) supplemented with 10% fetal bovine serum, 100 IU/mL penicillin, and 100 μg/mL streptomycin.

To evaluate the oestrogen-stimulated growth of these cells, they were released from monolayer using 0.25% trypsin/0.53 m M EDTA and suspended in oestrogen-free medium comprised of DMEM (high glucose, phenol red-free), supplemented with 10% charcoal-stripped bovine calf serum, 2 m M glutamine, 100 IU ml−1 penicillin, and 100  μg ml−1 streptomycin.