RT-PCR analysis of laminin chain expression.

Total RNA isolated from the adherent stromal layers of LTMCs was reverse transcribed, amplified with specific primer pairs for the human laminin α, β, and γ chains, and separated on a 2.0% agarose gel. The following specific amplification products were detected: a 357–base pair (bp) signal for the α1 chain, 368 bp for the α2 chain, 172 bp for the α3_A chain, 253 bp for the α3_B chain, 358 bp for the α4 chain, and 562 bp for the α5 chain; a 253-bp signal for the β1 chain and 337 bp for the β2 chain; a 283-bp signal for the γ1 chain and a 579-bp signal for the γ2 chain. No signals could be detected for the β3 chain and the γ3 chain, but positive controls with RNA from endothelial cells showed that the primer pairs for the β3 and the γ3 chain could amplify a 261-bp and a 556-bp signal, respectively. In addition, two 100-bp ladders (marker) and a β-actin control amplification are shown.