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Evaluation of cell proliferation and apoptosis

TT Tomomi Taguchi
YK Yoshio Kodera
KO Kazuhito Oba
TS Tatsuya Saito
YN Yuzuru Nakagawa
YK Yusuke Kawashima
MS Masayoshi Shichiri
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A10 cells seeded into 12-well cluster dishes (2.0 × 105 cells/well) were deprived of serum for 24 h and further incubated in the presence or absence of SBSN_HUMAN[225–237] or SBSN_HUMAN[243–259] for the indicated times. Cells were then trypsinized, and cell numbers measured using an EVE automatic cell counter (NanoEnTek Inc, Seoul, Korea)50.

HAoSMCs were cultured to subconfluency and serum deprived for 9 h in the presence or absence of SBSN_HUMAN[225–237] (10–6 M) or SBSN_HUMAN[243–259] (10–6 M). The terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay was performed using an ApopTag Fluorescein Direct In Situ Apoptosis Detection Kit (EMD Millipore, USA)51. DNA fragmentation was detected by laser scanning confocal microscopy using an LSM710 confocal microscope (Carl Zeiss, Jena, Germany).

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