FIGURE 3.

Knock-down of Physarum H1 modifies replication timing. A, experimental scheme for monitoring replicon firing. The cells were pulsed with BrdUrd for 1 or 3 h. Genomic DNA was isolated and digested either with HindIII or EcoRI (vertical arrows) prior to heat denaturation and pull-down using anti-BrdUrd antibody. The presence of replicons within the immunoprecipitated fraction was then analyzed by PCR and gel electrophoreses. B, H1 depletion accelerates firing of a late replicating origin. Comparison of timing of replication of Lav1.1 and Lav2.1 replicons in plasmodia untreated and treated with nonspecific siRNAs and H1 siRNAs.