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Cell culture and transfection

HK H. Kaur
JC J. Carvalho
ML M. Looso
PS P. Singh
RC R. Chennupati
JP J. Preussner
SG S. Günther
JA J. Albarrán-Juárez
DT D. Tischner
SC S. Classen
SO S. Offermanns
NW N. Wettschureck
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Isolated human aortal SMC (passage 1) were obtained from Cell Applications (Cat. No. 354-05a; San Diego, CA, USA) and Innoprot (Cat. No. P10456; Derio, Spain). Cells were thawed and immediately processed for expression analysis without further culture. Primary murine aortic SMC were isolated as described above and cultured in collagen I-coated (100 μg ml−1; Corning) 24-well plates for 7 days. Cells were transfected on day 0 with siRNAs directed against murine Gprc5b (Mm_Gprc5b_6; target sequence: 5′-TCGGGCCTACATGGAGAACAA-3′) or AllStars negative control siRNA using Lipofectamine RNAiMAX. All siRNAs were obtained from Qiagen.

Copyright and License information:  ©2017, The Author(s)
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