HEK-293 cells were divided into the miR-335-5p and pmirGLO empty vectors, miR-335-5p and pmirGLO-MAPK10-WT (GenePharma), miR-335-5p, and pmirGLO- MAPK10-MuT (GenePharma) co-transfection groups, respectively. Untreated cells were used as controls. Wild-type and mutant MAPK10 were synthesized by GeneChem (Shanghai, China) as follows: wild-type MAPK10, up 5ʹ-cATTTAACTTCTAGTTGCTCTTGCc-3ʹ and down 5ʹ-tcgagGCAAGAGCAACTAGAAGTTAAATgagct-3ʹ; and mutant MAPK10, up 5ʹ-cATTTAACTTCTAGTTGATATCGCc-3ʹ and down 5ʹ-tcgagGCGATATCAACTAGAAGTTAAATgagct-3ʹ. Cells were inoculated onto 96-well plates and cultured for 24 h. Luciferase activity was detected using a microplate reader. Renilla luciferase was used as an internal reference.
Pmirglo mapk10 wt
Manufactured by GenePharma
PmirGLO-MAPK10-WT is a lab equipment product that serves as a plasmid vector for gene expression. It contains the MAPK10 gene sequence in its wild-type form.
Automatically generated - may contain errors
4 protocols using pmirglo mapk10 wt
1
Investigating miR-335-5p and MAPK10 Interaction
2
Cell Cycle Analysis and Luciferase Assay
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Transfected cells in the logarithmic growth phase were inoculated onto a 6-well plate and cultured for 1 day. Cells were xed in 70% ethanol for 24 h and treated with propidium iodide and RNase provided with the kit. The cell cycle distribution was detected by ow cytometry.
Dual Luciferase Reporter Assay HEK-293 cells were divided into the miR-30a-3p and pmirGLO empty vectors, miR-335-5p and pmirGLO-MAPK10-WT (GenePharma), miR-335-5p, and pmirGLO-MAPK10-MuT (GenePharma) co-transfection groups, respectively. Untreated cells were used as controls. Wild-type and mutant MAPK10 were synthesized by GeneChem (Shanghai, China) as follows: wild-type MAPK10, up 5 -cATTTAACTTCTAGTTGCTCTTGCc-3 and down 5 -tcgagGCAAGAGCAACTAGAAGTTAAATgagct-3 ; and mutant MAPK10, up 5 -cATTTAACTTCTAGTTGATATCGCc-3 and down 5 -tcgagGCGATATCAACTAGAAGTTAAATgagct-3 . Cells were inoculated onto 96-well plates and cultured for 24 h. Luciferase activity was detected using a microplate reader. Renilla luciferase was used as an internal reference.
Dual Luciferase Reporter Assay HEK-293 cells were divided into the miR-30a-3p and pmirGLO empty vectors, miR-335-5p and pmirGLO-MAPK10-WT (GenePharma), miR-335-5p, and pmirGLO-MAPK10-MuT (GenePharma) co-transfection groups, respectively. Untreated cells were used as controls. Wild-type and mutant MAPK10 were synthesized by GeneChem (Shanghai, China) as follows: wild-type MAPK10, up 5 -cATTTAACTTCTAGTTGCTCTTGCc-3 and down 5 -tcgagGCAAGAGCAACTAGAAGTTAAATgagct-3 ; and mutant MAPK10, up 5 -cATTTAACTTCTAGTTGATATCGCc-3 and down 5 -tcgagGCGATATCAACTAGAAGTTAAATgagct-3 . Cells were inoculated onto 96-well plates and cultured for 24 h. Luciferase activity was detected using a microplate reader. Renilla luciferase was used as an internal reference.
3
Dual Luciferase Assay for miRNA-MAPK10 Interaction
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Transfected cells in the logarithmic growth phase were inoculated onto a 6-well plate and cultured for 1 day. Cells were xed in 70% ethanol for 24 h and treated with propidium iodide and RNase provided with the kit. The cell cycle distribution was detected by ow cytometry.
Dual Luciferase Reporter Assay HEK-293 cells were divided into the miR-335-5p and pmirGLO empty vectors, miR-335-5p and pmirGLO-MAPK10-WT (GenePharma), miR-335-5p, and pmirGLO-MAPK10-MuT (GenePharma) co-transfection groups, respectively. Untreated cells were used as controls. Wild-type and mutant MAPK10 were synthesized by GeneChem (Shanghai, China) as follows: wild-type MAPK10, up 5 -cATTTAACTTCTAGTTGCTCTTGCc-3 and down 5 -tcgagGCAAGAGCAACTAGAAGTTAAATgagct-3 ; and mutant MAPK10, up 5 -cATTTAACTTCTAGTTGATATCGCc-3 and down 5 -tcgagGCGATATCAACTAGAAGTTAAATgagct-3 . Cells were inoculated onto 96-well plates and cultured for 24 h. Luciferase activity was detected using a microplate reader. Renilla luciferase was used as an internal reference.
Dual Luciferase Reporter Assay HEK-293 cells were divided into the miR-335-5p and pmirGLO empty vectors, miR-335-5p and pmirGLO-MAPK10-WT (GenePharma), miR-335-5p, and pmirGLO-MAPK10-MuT (GenePharma) co-transfection groups, respectively. Untreated cells were used as controls. Wild-type and mutant MAPK10 were synthesized by GeneChem (Shanghai, China) as follows: wild-type MAPK10, up 5 -cATTTAACTTCTAGTTGCTCTTGCc-3 and down 5 -tcgagGCAAGAGCAACTAGAAGTTAAATgagct-3 ; and mutant MAPK10, up 5 -cATTTAACTTCTAGTTGATATCGCc-3 and down 5 -tcgagGCGATATCAACTAGAAGTTAAATgagct-3 . Cells were inoculated onto 96-well plates and cultured for 24 h. Luciferase activity was detected using a microplate reader. Renilla luciferase was used as an internal reference.
4
Luciferase Assay of MAPK10 Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HEK-293 cells were divided into the miR-30a-3p and pmirGLO empty vector, miR-335-5p and pmirGLO-MAPK10-WT (GenePharma), miR-335-5p and pmirGLO-MAPK10-MuT (GenePharma) co-transfection groups, respectively. Cells without treatment were used as control. The MAPK10 wild-type and mutant fragments were synthesized by Genechem, Shanghai, China, as follows: wild-type MAPK10, up 5 -cATTTAACTTCTAGTTGCTCTTGCc-3 and down 5 -tcgagGCAAGAGCAACTAGAAGTTAAATgagct-3 ; and mutant MAPK10, up 5 -cATTTAACTTCTAGTTGATATCGCc-3 and down 5 -tcgagGCGATATCAACTAGAAGTTAAATgagct-3 . These cells were inoculated onto the 96-well plates, and cultured for 24 h. The luciferase activity was detected by the microplate reader. Renilla was used as internal reference.
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