Figure 2. Mael Represses the Canonical Transcription of a Euchromatic Reporter in 42AB.

(A) RNA-seq, GRO-seq, piRNA, and H3K4me3, H3K9me3, Rhi, and HP1a ChIP-seq profiles for P{GSV6}42A18, a gfp transgene inserted in 42AB containing five tandem GAL4-binding upstream activating sequences with a core promoter derived from the Hsp70Bb gene (Toba et al., 1999). The gfp transgene contains an intron and canonical poly(A) sites in the 3′ UTR. Reads from mael^M391/r20 mutants are shown in red while wild-type controls are shown in black. Annotated transcription termination sites (red) are labeled. RNA- and GRO- seq data are for uniquely mapping reads, while piRNA data are from both uniquely and all mapping reads from the mean of three biological samples. ChIP-seq data are for uniquely mapping reads from the mean of two biological samples.

(B) Representative Western blots for GFP, Mael, or α-Tubulin (α-Tub) from ovaries with the genotype given below. GFP Western signal is the mean of three biological replicates normalized to α-Tub and is given in arbitrary units. p-values were measured using an unpaired, two-tailed t-test compared to w¹¹¹⁸; P{GSV6}42A18/+; +. Uncropped gel images can be found in Figure S2C.

(C) RNA-seq and piRNA profiles for P{GSV6}zip, which is identical in sequence to P{GSV6}42A18 but inserted into the first intron of the subtelomeric gene zip. RNA-seq data are for uniquely mapping reads, while all mapping piRNAs generated by the ping-pong and phased piRNA pathways are shown.

(D) Strategy to identify putative ping-pong and phased piRNAs, and to measure the probability of distances from the 5′ ends of putative ping-pong piRNAs to the 5′ ends of putative phased piRNAs mapping to P{GSV6}zip. Distance probability analyses are for all sequencing reads ≥24 nt. Red lines show non-parametric regression (LOWESS) of the data.

See also Figures S2–S4.