Fig. 3.

dsRNA knockdown assays reveal potential histone gene expression regulators. (A) Proteins enriched by a dNSAF ratio of 2 and associated with nucleic acids are knocked down by dsRNA over 72 h. cDNA is synthesized with iScript reverse transcriptase with a mixture of poly-A and random hexamer primers. H2A mRNA levels for each knockdown were calculated relative to the nonspecific dsRNA control using Tub84b as a reference gene. Compared with the nonspecific knockdown, CG11844, vig, and brahma show a negative effect on H2A mRNA expression. (B) LisH domain-containing proteins that are enriched in the HisC sample are knocked down by dsRNA as described in A. Compared with the nonspecific knockdown, Smu1, mahj, and mxc all showed a negative effect on the mRNA expression of H2A. Data plotted are averages and SDs from three separate knockdowns.