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3T3-L1 cells were obtained from the American Type Culture Collection and subcultured in DMEM containing 10% calf serum. 3T3-L1 cells were differentiated with the SVF as described above. Chinese hamster ovary (CHO) cells were obtained from the American Type Culture Collection and maintained in α-minimum essential medium (Invitrogen) containing 10% FBS or Opti-MEM (Invitrogen) without FBS. An siRNA specific for mouse BMP-3b (Silencer Select Pre-designed s66568) was purchased from Ambion (Austin, TX, USA). The targeted nucleotide sequence of the siRNA was CCACATGCCCTATATCCTT. Silencer Select Negative Control #1 (Ambion) was used as a control. For siRNA-mediated knockdown of 3T3-L1 cells, the cells were transfected with each siRNA (50 nm) using a Nucleofactor, plated at 1.25 × 105 cells cm−2 and then differentiated after 4 h. |
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Protocol tips |
3T3-L1 cells were obtained from the American Type Culture Collection and subcultured in DMEM containing 10% calf serum. 3T3-L1 cells were differentiated with the SVF as described above. Chinese hamster ovary (CHO) cells were obtained from the American Type Culture Collection and maintained in α-minimum essential medium (Invitrogen) containing 10% FBS or Opti-MEM (Invitrogen) without FBS. An siRNA specific for mouse BMP-3b (Silencer Select Pre-designed s66568) was purchased from Ambion (Austin, TX, USA). The targeted nucleotide sequence of the siRNA was CCACATGCCCTATATCCTT. Silencer Select Negative Control #1 (Ambion) was used as a control. For siRNA-mediated knockdown of 3T3-L1 cells, the cells were transfected with each siRNA (50 nm) using a Nucleofactor, plated at 1.25 × 105 cells cm−2 and then differentiated after 4 h. |
Publication protocol
3T3-L1 cells were obtained from the American Type Culture Collection and subcultured in DMEM containing 10% calf serum. 3T3-L1 cells were differentiated with the SVF as described above. Chinese hamster ovary (CHO) cells were obtained from the American Type Culture Collection and maintained in α-minimum essential medium (Invitrogen) containing 10% FBS or Opti-MEM (Invitrogen) without FBS. An siRNA specific for mouse BMP-3b (Silencer Select Pre-designed s66568) was purchased from Ambion (Austin, TX, USA). The targeted nucleotide sequence of the siRNA was CCACATGCCCTATATCCTT. Silencer Select Negative Control #1 (Ambion) was used as a control. For siRNA-mediated knockdown of 3T3-L1 cells, the cells were transfected with each siRNA (50 nm) using a Nucleofactor, plated at 1.25 × 105 cells cm−2 and then differentiated after 4 h.
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