Extended Data Fig. 6: Depletion of CDK-2 by the AID system and RNAi, and the dispensability of CDK-2 for homologue pairing, synapsis, and DSB induction.
From: Crossover patterning through condensation and coarsening of pro-crossover factors
a, Late prophase nuclei in aid::HA::cdk-2; sun-1p::tir1 hermaphrodites stained with anti-HA antibodies show incomplete depletion of CDK-2 by auxin treatment alone, while effective depletion by RNAi + auxin treatment. Arrowheads indicate the remaining CDK-2 signal. b, Early prophase nuclei in aid::HA::cdk-2; sun-1p::tir1 gonads stained for HIM-8, which marks the pairing center region of the X chromosome, and DNA. The robust pairing of HIM-8 foci was observed following depletion of CDK-2. c, Nuclei at mid-prophase stained for HTP-3 and SYP-1. Colocalization of these proteins reveals the formation of SCs along all axes in controls and hermaphrodites depleted of CDK-2. d, Nuclei at mid-prophase stained for RAD-51 and DNA, showing abundant DSB repair intermediates marked by RAD-51 following depletion of CDK-2. e, Dynamics of RAD-51 foci. Hermaphrodite gonads were divided into six zones of equal length spanning the premeiotic through the late pachytene region, as shown in the diagram. The number of RAD-51 foci in each nucleus is shown for each zone, with mean and s.d. values indicated by bars. The numbers of nuclei counted for zones 1, 2, 3, 4, 5, and 6 were as follows: for controls – 74, 113, 113, 127, 86, and 47, respectively; for CDK-2 depletion – 47, 46, 80, 82, 58, and 33, respectively. Data were derived from 2 animals for each condition, pooled from two independent replicates. More nuclei were present in the controls due to the effects of CDK-2 depletion on germline proliferation. ****P = 8.2210e-6 and 3.8779e-7, respectively. Two-tailed Mann-Whitney test. Scale bars in a-d: 5 µm. Source numerical data are available in source data.
