Bcl-2 siRNA (h)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 1.5 × 10^5/flask in 4 ml,	siRNA 5′ AACATCGCCCTGTGGATGACT-3′ Add diluted siRNA to HiPerFect Transfection Reagent to make a mixture   Incubate the mixture for 5–10 min at room temperature.  Add mixture to cells and incubate for 72 h

Upstream tips
Seed 1.5 × 10^5/flask in 4 ml,
Protocol tips
siRNA 5′ AACATCGCCCTGTGGATGACT-3′ Add diluted siRNA to HiPerFect Transfection Reagent to make a mixture   Incubate the mixture for 5–10 min at room temperature.  Add mixture to cells and incubate for 72 h

Publication protocol

Exponentially growing untreated MCF-7 and MDA-MB-231 cells were collected and plated (2 and 1.5 × 105/flask in 4 ml, respectively) 24 hours before transfection. Plated cells were transfected with either Bcl-2 siRNA or control siRNA (50 nmol/l). siRNA sequences targeting Bcl-2 and nonsilencing control siRNA 5′-AACATCGCCCTGTGGATGACT-3′ and 5′-AATTCTCCGAACGTGTCACGT-3′, respectively.17 Beclin-1 siRNA and ATG8 siRNA22 were used. The siRNAs were dissolved in sterile buffer provided by the manufacturer (all from Qiagen Inc, Valencia, CA, USA). On the day of transfection, 1.5 µg of siRNA was mixed with HiPerFect transfection reagent according to the manufacturer's instructions (Qiagen) and added to the cells in each well.

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Manufacturer protocol

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