BECN1 siRNA (m)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	siRNA concentartion 30 pM. Incubate for 30 minutes at room temperature with siRNA and reagent mixture. Add mixture to cells and incuabte for 4h. Change medium and incubate for another 20h

Protocol tips
siRNA concentartion 30 pM. Incubate for 30 minutes at room temperature with siRNA and reagent mixture. Add mixture to cells and incuabte for 4h. Change medium and incubate for another 20h

Publication protocol

BV-2 cells were seeded into 6-well plates to grow about 80% confluent. The next day, individual targeted siRNA and scrambled siRNA (30 pM) were mixed with lipofectamine 2000 (2 µl) in 100 µl opti-MEM (Life technologies, 31985062). After 30 min incubation at room temperature, mixed liquids were dropped into cell culture medium (serum free) and were incubated for 4 h. Then the medium was changed to 10% FBS-containing medium for 20 h incubation. The transfected cells were then ready for the following experiments.

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