Fig. 6.
SNAP suppressed LPS-induced NGF mRNA expression. Cells were stimulated with LPS in the presence of SNAP, dibutyryl-cGMP, and NMMA for 12 h. Then, total RNA was extracted, and cDNA was made for the performance of RT-PCR. To more accurately quantify mRNA levels, NGF cDNA amplified by nonsaturated PCR cycles (n = 20) was separated in an acrylamide gel and stained with the sensitive dye SYBR Green I (with a detectable range of 5–1000 pg DNA), and the fluorescence intensity of the products was quantified using a fluorescence image analyzer (Fuji FLA-2000; n = 4). The values were normalized with those of PCR products amplified from β-actin cDNA (15 PCR cycles).