Stalk cells were isolated from 24 h fruiting bodies by pipetting vigorously in pronase/EDTA, followed by deposition on 50 µm nylon mesh and extensive washing with KK2. The stalks retained on the mesh were then collected, placed on top of 5 ml 25% Percoll in KK2 and centrifuged at 10,000g for 5 minutes at 4ºC. Stalks were collected from the top layer, washed with 5 volumes of KK2 and pelleted at 4,000 x g for 3 minutes. Dictyostelium lacteum amoebas were co-cultured with Klebsiella aerogenes on lactose-peptone agar. RNeasy kit (QIAGEN) was used for extraction. Libraries were prepared with Illumina Truseq stranded mRNA kit.
Experimental Factor: cell type
Stalk
Experimental Factor: time
24