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. 1999 Dec;155(6):2043–2055. doi: 10.1016/S0002-9440(10)65523-X

Figure 1.

Figure 1.

Suppression subtractive hybridization screening. A: PCR-based cDNA synthesis from 1 μg HEVEC total RNA. Lanes 1 and 2: Hybridization analysis of HEVEC cDNA integrity with human PAPS synthetase (lane 1) or E-selectin (lane 2) probes. Lanes 3 and 4: Ethidium bromide staining of HEVEC cDNAs before (lane 3) and after (lane 4) Rsa-1 digestion. B: PCR analysis of the efficiency of PAPS synthetase cDNA subtraction. The abundance of PAPS synthetase cDNA fragments in the unsubtracted HEVEC and the subtracted HEVEC-HUVEC cDNA populations was compared by performing PCR amplification for the indicated number of cycles. C: Southern blot analysis of the isolated cDNAs with HEVEC-HUVEC and HUVEC-HEVEC subtracted probes. The isolated cDNAs were released from plasmid DNA by digestion with EcoRI and XhoI, separated on a 1% agarose gel, and transferred onto nylon filters. Duplicate filters were then hybridized with HEVEC-HUVEC and HUVEC-HEVEC subtracted probes. Lanes 1, 3, and 4: Differentially expressed cDNAs encoding thrombospondin-1, hevin, and mac25/IGFBP-rP1, respectively. Lane 2: cDNA not detected by the subtracted probes in the secondary screen. Lane 5: cDNA similarly detected with the two probes.

HHS Vulnerability Disclosure