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. 2011 Sep 20;108(40):16681–16686. doi: 10.1073/pnas.1114209108

Fig. 2.

Fig. 2.

DAFC-34B exhibits two stages of replication initiation, the second of which is transcription-independent. Genomic DNA was isolated from hand-sorted staged egg chambers, and DNA copy levels were quantified by qPCR compared with a nonamplified locus at 62C5. Error bars show SD for triplicate reactions. Genomic position is shown on the x-axis (13380 is Chr2L:13,380,000). (A) Gene amplification is not first observed at stage 9, despite expression of Vm34Ca. (B) Replication initiation occurs in stage 10B to reach threefold amplification, followed by a second round of replication initiation at stage 13. (C and D) Whole ovaries were in vitro- cultured with α-amanitin for 5 h, after which staged egg chambers were hand-dissected and analyzed as in A and B. qPCR quantification of stage 10 and stage 13 are shown for DAFC-34B and DAFC-62D. (C) At stage 10, α-amanitin treatment has no effect on amplification at either amplicon. (D) At stage 13, α-amanitin treatment specifically inhibits late amplification at DAFC-62D, but has no effect at DAFC-34B.