| Id |
Subject |
Object |
Predicate |
Lexical cue |
| T55 |
0-48 |
Sentence |
denotes |
Transient transfection and western blot analysis |
| T56 |
49-244 |
Sentence |
denotes |
293FT cells were seeded onto 6-cm dishes 24 hours before transient transfection using X-tremeGENE HP DNA transfection reagent (Roche, Basel, Switzerland) according to the manufacturer’s protocol. |
| T57 |
245-385 |
Sentence |
denotes |
At 24 hours post-transfection, cells were harvested, spun down by centrifugation and washed with cold phosphate buffered saline (PBS) twice. |
| T58 |
386-464 |
Sentence |
denotes |
Cells were then resuspended in 2× Laemmli sample buffer, boiled and sonicated. |
| T59 |
465-667 |
Sentence |
denotes |
Clarified supernatant containing the protein of interest was obtained by spinning down the cell lysate at 13,000 rpm at 4 °C to remove the cell debris and further analysed by western blot (WB) analysis. |
| T60 |
668-861 |
Sentence |
denotes |
Equal amounts of total cell lysates were loaded per lane and resolved using electrophoresis on SDS-PAGE gels and transferred onto nitrocellulose membrane (Bio-Rad, Hercules, CA, United States). |
| T61 |
862-1049 |
Sentence |
denotes |
The membrane was blocked in 5% skimmed milk in Tris-buffered saline with 0.05% Tween 20 (TBST) for 1 hour at room temperature (RT) and incubated with primary antibodies at 4 °C overnight. |
| T62 |
1050-1218 |
Sentence |
denotes |
After the membrane was washed with TBST, it was incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody (Thermo Fisher Scientific) at RT for 1 hour. |
| T63 |
1219-1408 |
Sentence |
denotes |
The membrane was then washed with TBST again and bound antibodies visualised with enhanced chemiluminescence substrate (Thermo Fisher Scientific) using ChemiDoc MP Imaging System (Bio-Rad). |
