Wwtr1 siRNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	siRNA concentartion-25 pmol Incubate Lipofectamine reagent:siRNA mixture for 5 min and add to cells. Add mixture to cells for 24h and later change medium. Incubate cells for 1–3 days at 37°C.

Protocol tips
siRNA concentartion-25 pmol Incubate Lipofectamine reagent:siRNA mixture for 5 min and add to cells. Add mixture to cells for 24h and later change medium. Incubate cells for 1–3 days at 37°C.

Publication protocol

Wild‐type (WT) TAZ, TAZ S89A, YAP S127A, or WT YAP was subcloned into a pMSCV‐IRES‐eGFP retroviral expression backbone (Addgene Plasmids 24809, 24815, 17791 and 17790) creating pMSCV‐3xFlag TAZ‐IRES‐eGFP and pMSCV‐3xFlag‐TAZ S89A‐IRES‐eGFP 42. Empty vector was negative control. Retroviruses were packaged in HEK293T cells using standard methods. Medium was changed 1 hour before transfection/transduction. Retroviral suspension diluted 1:4 with polybrene (4 μg/mL) was added for 6 h, before changing medium.

Taz small interfering RNA (siRNA) (Ambion (http://www.ambion.com/), Foster City, California, USA, s97145) and Yap siRNA (Ambion, s202423) were used as per manufacturer's instructions. For plated satellite cells, 25 pmol of siRNA with Lipofectamine RNAiMax (ThermoFisher Scientific) was added to each well for either 6 hours (satellite cells) or 24 hours (C2C12) before medium was changed.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Wwtr1 siRNA below.

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