siRNA FOXS1

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio).  Incubate for 5 minutes at room temperature and add to cells.  Incubate cells for 1–3 days at 37°C.

Protocol tips
Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio).  Incubate for 5 minutes at room temperature and add to cells.  Incubate cells for 1–3 days at 37°C.

Publication protocol

Predesigned GLI1 siRNA (Villegas et al., 2014) and nontargeting control siRNA (SIC001) were purchased from Sigma‐Aldrich (St. Louis, MO, USA), while siRNA targeting FOXS1 (sc‐75023) were purchased from Santa Cruz Biotechnologies (Dallas, TX, USA). Cells were plated in 6‐ or 24‐well plates at 50–70% confluency, and transfections were performed with Lipofectamine RNAimax (Invitrogen, Carlsbad, CA, USA) following the protocol provided by the manufacturer. Rh36 cells were transfected with expression constructs for GLI1 (Shimokawa et al., 2013), GLI1‐701G (Shimokawa et al., 2013), and 8 control pCMV‐based constructs or siRNA that did not cause significant change in GLI1 mRNA level. Transfections were performed using Lipofectamine LTX (Invitrogen) following manufacturer's instructions.

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Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for siRNA FOXS1 below.

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