MiR-Inhibition Transfection Protocol

The miR-inhibition was performed with the addition of 50 nM of miR-294-3p-, miR-301b-3p-, or miR-410-3p-specific inhibitors (cat 4464084) or the negative control (cat 4464076) (miRVana, Ambion, Thermo Scientific) mixed with 6 μL FuGene Reagent Transfection (Roche, Promega, Madson, WI, USA), which was incubated for 20 min at room temperature following our previously described protocol [62 (link)] in 500 μL of serum-free RPMI 1640 medium (LGC Biotecnologia, São Paulo, SP, Brazil) [27 (link)]. After 24 h of transfection, the cells were infected as described above.

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Acuña S.M., Zanatta J.M., de Almeida Bento C., Floeter-Winter L.M, & Muxel S.M. (2022). miR-294 and miR-410 Negatively Regulate Tnfa, Arginine Transporter Cat1/2, and Nos2 mRNAs in Murine Macrophages Infected with Leishmania amazonensis. Non-Coding RNA, 8(1), 17.

Publication 2022

RPMI 1640 medium

Cells Fugene 6 Inhibition Inhibitors Nm 294 Promega Serum Transfection

Corresponding Organization : Universidade de São Paulo

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Variable analysis

independent variables

Concentration of miR-294-3p-, miR-301b-3p-, or miR-410-3p-specific inhibitors (50 nM)

dependent variables

Not explicitly mentioned

control variables

Negative control (cat 4464076) (miRVana, Ambion, Thermo Scientific)
FuGene Reagent Transfection (6 μL)
Incubation time (20 min at room temperature)
Serum-free RPMI 1640 medium (500 μL)

controls

Negative control (cat 4464076) (miRVana, Ambion, Thermo Scientific)

Annotations

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