RT-PCR of cytokine receptors, SAA and SAF-1 transcripts in non-differentiated and
differentiated hMSCs: hMSCs were kept in either expansion or osteogenic medium
and stimulated with different cytokines (10 ng/ml) for 24 h. A:
Human cytokine receptor transcripts as well as (B)
SAA and SAF transcripts were amplified
using specific oligonucleotide primers (Supplement, Table I). The PCR products were separated on
1.5% agarose gels. NS, non-stimulated; P, positive control: RNA was isolated
from HUH-7 cells for SAF-1, SAA, cytokine receptors except IL-1R2, where RNA
from THP-1 cells was used; N1, negative control— RNA template: negative
controls were done for all samples; N2, negative control—water template.
To ensure that equal amount of cDNA was used as a template RT-PCR for GAPDH was
made as a control. One representative experiment out of three is shown.