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Highly Precise and Developmentally Programmed Genome Assembly in Paramecium Requires Ligase IV–Dependent End Joining

Figure 2

Induction of LIG4 and XRCC4 transcription during autogamy.

A. Northern blot hybridization of total RNA extracted from strain 51 during an autogamy time-course in standard growth medium. The blot was hybridized successively with LIG4, XRCC4 (see maps in Figure 4A) and 17S rDNA probes. The histograms show the progression of autogamy, with the different cellular stages diagrammed on top. Bottom panel shows the LMPCR-mediated detection of DSBs at the left boundary of IES sm19-576 (broken MAC ends). V: vegetative cells. The following time-points are indicated in hrs. Time 0 corresponds to 50% of the cells harboring a fragmented old MAC. B. Quantification of mRNA levels from the blots shown in A. For PiggyMac, values were calculated from a previous hybridization of the same blot [12]. 17S rRNA signal was used for normalization. Y-axes are in arbitrary units.

Figure 2