Lentivirally shRNA transduced U87MG, NCH421k and NCH466 cells were cultured for 16 h (U87MG, 20% confluence) or 7 days (NCH421k and NCH644) prior to analysis.
To produce cells stably expressing AC2 constructs, HEK-293 cells were plated at ~60% confluence 1 day prior to transfection with 2 μg of constructs.
Thompson, Lance M., 21, Upland, Ind .Thompson, Nils G., 19, Confluence, Pa.
(CBD common bile duct, LHD left hepatic duct, RHD right hepatic duct, B1l confluence of the left caudate branch, B1r: confluence of the right caudate branch).
The recombinant SCARB2 protein was produced using Sf9 cells by infecting 1 liter of cells (2 × 106/mL confluence) with 10 mL of P3 viral stock that was obtained following the manufacturer's instructions (Invitrogen).
These shared features include the presence of four-bounding membranes and, with some exceptions [29], confluence of the outermost plastid membrane with the nuclear envelope [30].
Lymphoblastoid cell suspension cell cultures were collected at 6 × 10 confluence and processed for protein extraction.
Lymphoblastoid cell suspension cell cultures were collected at 6 × 10 confluence and processed for RNA extraction.
Additionally, we performed the whole-genome DNA profiling of 80MLa culture also at second (p2) confluence (see Additional File 2) and of 81BPG culture at second (p2) and third (p3) confluences.
When combined with miR-181a inhibitor, 50 or 100 μM of miR-181a inhibitor was transfectedinto cells (~70% confluence in 12-well plates ) 12 hours before bufalin treatment.
In a separate experiment, we also assessed the methylation levels in dividing EpH4 cells, grown at ~70% confluence (12 replicates).
