Vybrant™ MTT Cell Proliferation Assay Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cells were pretreated with NC or miR-133a mimics respectively.	- After 48h , cells wereassayed for apoptosis. - Cells were treated with YO-PRO®-1 stock solution and PI stock solution and incubated for for 20–30 minutes on ice.

Upstream tips
- Cells were pretreated with NC or miR-133a mimics respectively.
Protocol tips
- After 48h , cells wereassayed for apoptosis. - Cells were treated with YO-PRO®-1 stock solution and PI stock solution and incubated for for 20–30 minutes on ice.

Publication protocol

Osteosarcoma MG63 or U2OS cells were transfected with NC or miR-133a mimics respectively. At 48 h post transfection, spent cell culture medium was replaced with serum free DMEM and placed in 1% oxygen incubator. In the indicated time periods post serum deprivation, cells were harvested, washed, resuspended in the staining buffer, and examined with Vybrant Apoptosis Assay kit (Invitrogen). Stained cells were detected by FACSCalibur and data were analyzed with CellQuest software (both from Becton Dickinson). The Annexin V-positive cells were regarded as apoptotic cells.

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