SmGMTM- 2 Smooth Muscle Cell Growth Medium -2 BulletKitTM

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Human primary Coronary Artery Smooth Muscle Cells (HCASMC, Lonza #CC-2583 Lot 200,212 and Cell Applications # 350-05a Lot 1508) were cultured in Smooth Muscle Growth Medium-2 including hEGF, insulin, hFGF-B and FBS, but without antibiotics (Lonza, #CC-3182).	Cells were changed to medium with supplements at 18 h post-transfection and cultured for an additional 48 h.

Protocol tips
Human primary Coronary Artery Smooth Muscle Cells (HCASMC, Lonza #CC-2583 Lot 200,212 and Cell Applications # 350-05a Lot 1508) were cultured in Smooth Muscle Growth Medium-2 including hEGF, insulin, hFGF-B and FBS, but without antibiotics (Lonza, #CC-3182).
Downstream tips
Cells were changed to medium with supplements at 18 h post-transfection and cultured for an additional 48 h.

Publication protocol

Human primary Coronary Artery Smooth Muscle Cells (HCASMC, Lonza #CC-2583 Lot 200,212 and Cell Applications # 350-05a Lot 1508) were cultured in Smooth Muscle Growth Medium-2 including hEGF, insulin, hFGF-B and FBS, but without antibiotics (Lonza, #CC-3182). For RNA-Seq studies donor-pooled HCASMC were transfected with 300 nM TCF21 Trilencer-27 Human siRNA (OriGene #SR304753C) or Trilencer-27 Universal Scrambled Negative Control siRNA (OriGene #SR30004) at 80% confluence using the Amaxa Basic Nucleofector Kit for Primary Mammalian Smooth Muscle Cells (Lonza #VPI-1004) at a density of 1 × 106 cells per 100 μL sample using Nucleofector Program U-025. Cells were changed to medium with supplements at 18 h post-transfection and cultured for an additional 48 h.

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