Id |
Subject |
Object |
Predicate |
Lexical cue |
T9960 |
0-27 |
Sentence |
denotes |
Constructs and transfection |
T181 |
0-27 |
Sentence |
denotes |
Constructs and transfection |
T9961 |
28-135 |
Sentence |
denotes |
The complementary DNA (cDNA) of the mouse Sharpin gene was cloned and amplified from RAW264.7 RNA extracts. |
T182 |
28-135 |
Sentence |
denotes |
The complementary DNA (cDNA) of the mouse Sharpin gene was cloned and amplified from RAW264.7 RNA extracts. |
T183 |
136-265 |
Sentence |
denotes |
The primer sequences were forward, 5′-CC ATG GCG ATG TCG CCG CCC GCC GGC GGT; reverse, 5′- AAG CTT CTA GGT GGA AGC TGC AGC AAG A. |
T9962 |
136-329 |
Sentence |
denotes |
The primer sequences were forward, 5′-CC ATG GCG ATG TCG CCG CCC GCC GGC GGT; reverse, 5′- AAG CTT CTA GGT GGA AGC TGC AGC AAG A. The Sharpin cDNA was cloned into expression vector pFLAG-CMV-2. |
T184 |
266-329 |
Sentence |
denotes |
The Sharpin cDNA was cloned into expression vector pFLAG-CMV-2. |
T9963 |
330-419 |
Sentence |
denotes |
Murine fibroblasts and macrophages were were used to express recombinant SHARPIN protein. |
T185 |
330-419 |
Sentence |
denotes |
Murine fibroblasts and macrophages were were used to express recombinant SHARPIN protein. |
T9964 |
420-500 |
Sentence |
denotes |
Cells (2×104) were seeded in 96-well treated plates the day before transfection. |
T186 |
420-500 |
Sentence |
denotes |
Cells (2×104) were seeded in 96-well treated plates the day before transfection. |
T9965 |
501-607 |
Sentence |
denotes |
After overnight incubation, 200 ng pFLAG-SHARPIN plasmids were transfected with 0.5 ul Lipofectamine 2000. |
T187 |
501-607 |
Sentence |
denotes |
After overnight incubation, 200 ng pFLAG-SHARPIN plasmids were transfected with 0.5 ul Lipofectamine 2000. |
T9966 |
608-671 |
Sentence |
denotes |
24 hours later, cells were incubated with fresh culture medium. |
T188 |
608-671 |
Sentence |
denotes |
24 hours later, cells were incubated with fresh culture medium. |
T9967 |
672-782 |
Sentence |
denotes |
After another 24 hours, cells were lysed to confirm the FLAG-SHARPIN expression by immunoblots with anti-FLAG. |
T189 |
672-782 |
Sentence |
denotes |
After another 24 hours, cells were lysed to confirm the FLAG-SHARPIN expression by immunoblots with anti-FLAG. |
T9968 |
783-918 |
Sentence |
denotes |
Cells with no transfection and transfected with empty vector pFLAG-CMV-2 were used as negative control in all transfection experiments. |
T190 |
783-918 |
Sentence |
denotes |
Cells with no transfection and transfected with empty vector pFLAG-CMV-2 were used as negative control in all transfection experiments. |