Lipofectamine 3000 and P3000™ Enhancer Reagent (Thermo Fisher
Scientific Inc) were used for transient plasmid transfections according to
manufacturer’s protocol and the cells were cultured for 24 h. SiRNA
silencing was performed using 100 nM siRNA and Lipofectamine® RNAiMAX
Reagent (Thermo Fisher Scientific Inc) according to manufacturer’s
protocol and cells were cultured for 3 days. The siRNAs targeting human SHANK1
and SHANK3 were SMARTpool ON-TARGETplus purchased from Dharmacon (SHANK1 Cat.
No. L-031945-02-0005; SHANK3 Cat. No. L-024645-00-0005). SiRNAs used as controls
were Allstars negative control siRNA (Qiagen, Cat. No. 1027281) and
ON-TARGETplus Non-targeting Pool (Dharmacon, Cat. No. D-001810-10-05). Rescue
experiments consisted of plasmid transfections 72 h after siRNA silencing of the
protein of interest.
Scientific Inc) were used for transient plasmid transfections according to
manufacturer’s protocol and the cells were cultured for 24 h. SiRNA
silencing was performed using 100 nM siRNA and Lipofectamine® RNAiMAX
Reagent (Thermo Fisher Scientific Inc) according to manufacturer’s
protocol and cells were cultured for 3 days. The siRNAs targeting human SHANK1
and SHANK3 were SMARTpool ON-TARGETplus purchased from Dharmacon (SHANK1 Cat.
No. L-031945-02-0005; SHANK3 Cat. No. L-024645-00-0005). SiRNAs used as controls
were Allstars negative control siRNA (Qiagen, Cat. No. 1027281) and
ON-TARGETplus Non-targeting Pool (Dharmacon, Cat. No. D-001810-10-05). Rescue
experiments consisted of plasmid transfections 72 h after siRNA silencing of the
protein of interest.