For depletion of PINK1 in HEK 293 cells, the following siRNAs were tested: PINK1 siRNA-1 (1199, Ambion, Grand Island, NY, U.S.A.), 5′-GGAGAUCCAGGCAAUUUUUTT-3′ and PINK1 siRNA-2, 5′-GACGCUGU UCCUCGUUAUGAA-3′ (S100287931, Qiagen, Venlo, Netherlands). Scrambled siRNAs with no known mammalian homology (non-targeting siRNA #1 (Ambion) and #2 (Santa Cruz Biotechnology, Dallas, TX, U.S.A.)) were used as negative controls. HEK cells were transfected with the siRNAs using the Mirus TransIT-TKO® reagent (Mirus, Madison, WI, U.S.A.) according to the manufacturer’s manual and then harvested after 48 h. Control vector-transfected cells were used as controls for all the experiments.
Mirus transit tko reagent
Manufactured by Mirus Bio
Sourced in United States
Mirus TransIT-TKO® reagent is a proprietary lipid-based transfection reagent designed for efficient delivery of DNA, RNA, and other macromolecules into a variety of cell types.
Automatically generated - may contain errors
Lab products found in correlation
Non targeting sirna 1, by Thermo Fisher Scientific (1 mentions)
Signalsilence jnk sirna, by Cell Signaling Technology (1 mentions)
Scrambled duplex control sirna, by Cell Signaling Technology (1 mentions)
Signalsilence p38 mapk sirna, by Cell Signaling Technology (1 mentions)
Signalsilence control sirna, by Cell Signaling Technology (1 mentions)
3 protocols using mirus transit tko reagent
1
Depletion of PINK1 in HEK 293 cells
2
Silencing MAPK signaling in myocytes
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The expressions of MAPKs were silenced by small RNAs interference technique in this study. SignalSilence p38 MAPK siRNA (#6564, Cell Signaling Tech), SignalSilence p44/42 (Erk1/2) siRNA (#6560, Cell Signaling Tech) and SignalSilence JNK siRNA (#6232, Cell Signaling Tech) were used to transfect the cells. The scrambled duplex control siRNA (#6568, Cell Signaling Tech) was used as a negative control. The siRNAs were introduced to cultured myocytes by transfection with Mirus TransIT-TKO reagent (Mirus) in six-well culturing dishes according to the instructions provided by the manufacturer. The final concentrations of siRNAs were 100 nmol/L.
3
Silencing MKK3 and MKK6 in RAECs
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In this study, MKK3 and MKK6 were silenced in RAECs by using an siRNA transfection technique. SignalSilence MKK3 siRNA (Cat.6294S; Cell Signaling Technology, Danvers, MA) was used to knock down mkk3. The specific sequence of siRNA against mkk6 was: 3′‐CUACAGUAGUGAAGAGAUUTT‐3′, which was designed and synthesized by GenePharma Co, Ltd (Shanghai, China). SignalSilence Control siRNA (Cat.6568S; Cell Signaling Technology) was used as control. Cultured RAECs (at 70% confluence) were transfected with the above siRNAs with Mirus TransIT‐TKO reagent (Mirus Bio LLC, Madison, WI) for 48 hours per the manufacturer's instructions.
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