PICK1 siRNA

Protocol tips

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	After four hours of incubation, add serum to cells. After 24 hours replace Opti-MEM with DMEM and repeat the same process thrice to increase the transfection efficiency.

Protocol tips
After four hours of incubation, add serum to cells. After 24 hours replace Opti-MEM with DMEM and repeat the same process thrice to increase the transfection efficiency.

Publication protocol

siRNAs were transfected into astrocytes using Lipofectamine RNAiMax: astrocytes were transferred into serum-free Opti-MEM (Invitrogen) and transfected with siRNAs, as published previously (Benfenati et al., 2011). After 4 h of incubation, serum was added to transfected cells, obtaining a final concentration of 10% FBS. On the following day, Opti-MEM was replaced with DMEM-based cultivation medium, as described above. To increase the transfection efficiency, this procedure was repeated 3 days after the initial transfection. siRNAs directed against Arp3 (Korobova and Svitkina, 2008) and WAVE2 (Danson et al., 2007) were purchased from Dharmacon (Lafayette, CO, US). The N-WASP-specific siRNA, based on a previously published shRNA (Kovacs et al., 2011), was purchased from Sigma-Aldrich. The PICK1-specific siRNA is based on a validated shRNA sequence (Citri et al., 2010) and was purchased from Sigma-Aldrich. The unspecific siRNA (5′-AGGUAGUGUAAUCGCCUUGTT-3′) was purchased from Eurofins MWG (Regensburg, Germany).

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for PICK1 siRNA below.

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