Journal:

Article Title: Disruption of actin cytoskeleton mediates loss of tensile stress induced early phenotypic modulation of vascular smooth muscle cells in organ culture

doi: 10.1016/j.yexmp.2009.10.006

Figure Lengend Snippet: Role of actin cytoskeleton dynamics in organ culture (OC)-induced dedifferentiation of VSMC. (A) the downregulation of VSMC-specific genes can be partially blocked by Jasplakinolide (Jasp), an actin polymerization inducer. (B) The actin cytoskeleton dynamics in organ culture was measured by the G-/F-actin ratio using western blot analysis and the intensity of each band was quantified by the Photoshop software. * and ** indicate p<0.05 and p<0.01 vs fresh group; ### indicates p<0.001 vs OC (24 h) group.

Article Snippet: Real-time PCR was performed using the StepOnePlus system (Applied Biosystems) in the presence of SYBR Green. snRNA U6 was used as the internal control, and all PCR primers were designed to cover at least 2 exons ( ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Sequence Amplicon size (bp) GenBank accession # U6 Forward 5’-CTCGCTTCGGCAGCACATATACTA-3’ 92 {"type":"entrez-nucleotide","attrs":{"text":"NR_002752","term_id":"366393295","term_text":"NR_002752"}} NR_002752 Reverse 5’-CGCTTCACGAATTTGCGTGTCATC-3’ SMA Forward 5’-ATCTTTTCCATGTCGTCCCAGTTG-3’ 310 {"type":"entrez-nucleotide","attrs":{"text":"NM_007392","term_id":"440309867","term_text":"NM_007392"}} NM_007392 Reverse 5’-GAGAAGCCCAGCCAGTCG-3’ SM22 Forward 5’-TCCTTCCAGTCCACAAACGACCAA-3’ 92 {"type":"entrez-nucleotide","attrs":{"text":"NM_011526","term_id":"291045204","term_text":"NM_011526"}} NM_011526 Reverse 5’-TTTGGACTGCACTTCTCGGCTCAT-3’ SM22 Forward 5’-TCCTTCCAGCCCACAAACGACCAA-3’ 92 {"type":"entrez-nucleotide","attrs":{"text":"NM_031549","term_id":"126506302","term_text":"NM_031549"}} NM_031549 Reverse 5’-TTTGGACTGCACTTCTCGGCTCAT-3’ SM-MHC Forward 5’-AACGCCCTCAAGAGCAAACTCAGA-3’ 161 {"type":"entrez-nucleotide","attrs":{"text":"NM_013607","term_id":"241982715","term_text":"NM_013607"}} NM_013607 Reverse 5’-TCCCGAGCGTCCATTTCTTCTTCA-3’ SRF Forward 5’-AAAGAAACCGTGAAAAGGCAACA-3’ 229 {"type":"entrez-nucleotide","attrs":{"text":"NM_020493","term_id":"133892959","term_text":"NM_020493"}} NM_020493 Reverse 5’-CTGACAGCACCGTCTGGGACCGT-3’ Myocd Forward 5’-CAGTGAAGCAGCAAATGACTCGG-3’ 230 {"type":"entrez-nucleotide","attrs":{"text":"NM_145136","term_id":"154240674","term_text":"NM_145136"}} NM_145136 Reverse 5’-GTCGTTGGCGTAGTGATCGAAGG-3’ Open in a separate window Sequences of primers used for real-time PCR Analysis of actin cytoskeleton dynamics The G/F-actin ratio in vivo assay kit (Cytoskeleton, Denver, CO) was used to measure the actin cytoskeleton dynamics in the aorta cultured for 24 hours with and without 0.3 µM Jasp based on the manufacturer’s protocol.

Techniques: Organ Culture, Western Blot, Software