Extended Data Figure 4 |. Complementation experiments performed with small DNA or antisense sRNA do not promote repair.

a, BELD quantitation of breaks on the forward (coding) strand of the CASP16P Pause2 termination site in BRCA1-depleted HeLa cells complemented with either ActB- or Casp16p-sdRNA, showing that only Casp16p-sdRNA can prevent accumulation of breaks at CASP16P Pause2. A representative experiment is shown, and the histograms depict the average values of the qPCR replicates ± s.d. b-c, DNA damage quantitation using γ-H2AX ChIP qPCR analyses performed in complementation experiments at the ACTB and CASP16P pause sites and at the CCNB1 CoTC. ChIP data are represented as the average fold change compared to an undepleted relevant control. Complementation experiments were performed in (b) BRCA1- or (c) Dicer-depleted HeLa cells reconstituted with the 3’ -non polymerizable sense sdRNA (3’-blocked ActB-sdRNA)(b-c), or ActB or Casp16p antisense (AS) sdRNA (b-c) or the corresponding sense or antisense (AS) sdRNA DNA sequence (b) (n = 2–6 biological replicates for b and n=2–6 for c). Data in b and c, were analyzed by Two-way Anova with post-hoc Tukey and compared to the relevant control cells.