Skip to main content
. 2019 Jan 16;8:e40806. doi: 10.7554/eLife.40806

Figure 3. Ova interacts with the heterochromatin machinery.

(a) Y2H assay for protein interaction between Ova and proteins as indicated. (b–c) Western blots showing reciprocal co-IP between Ova and HP1a, and between Ova and dLsd1. The RFP-HP1a transgene was driven by the endogenous promoter. The dLsd1-GFP transgene was driven by a ubiquitous promoter. The Flag-ova transgene was driven by nos-GAL4. (d) Heat map displaying steady state mRNA levels as reads per million (rpm) for the top 60 detected transposons in nosGAL4 driven ova-RNAi, EGFP-RNAi, and w1118 ovaries. The average of three replicates is shown. The most upregulated transposons are highlighted in bold. (e) Correlation scatter plot of log10 transposon mRNA-seq reads between ova GLKD and dLsd1 GLKD ovaries. R = 0.9463, p<2.2×10−16 by Pearson’s correlation coefficient. The most upregulated transposons in both genotypes are highlighted in red dots.

Figure 3.

Figure 3—figure supplement 1. Protein interaction mapping among components of heterochromatin machinery by Y2H assay.

Figure 3—figure supplement 1.

Y2H assay to test the interaction between proteins that associated with Piwi/piRNAs complexes and proteins in the heterochromatin machinery. SD-WL, non-selective medium; SD-WLH and SD-WLHA, selective media. Ф, empty vector.

Figure 3—figure supplement 2. Ova acts as a suppressor of position-effect variegation (PEV).

Figure 3—figure supplement 2.

(a) Photographs showing representative eye pigments of adult females bearing X chromosome wm4h reporter plus wild-type and ova heterozygous alleles. (b) Photographs showing representative eye pigments of adult females bearing fourth chromosome PEV reporters plus heterozygous or trans-heterozygous of ova mutant alleles. (c) Quantitative measure of eye pigment levels of indicated genotypes by a spectrophotometer. Values are mean ± SEM.; n > 5. P values by two-tailed Student’s t-test.

Figure 3—figure supplement 3. Ova is not required for piRNA biogenesis.

Figure 3—figure supplement 3.

(a) A scatter plot shows gene expression (rpkm, reads per kilobase per million sequenced reads) comparison of protein-coding genes between w1118 and ova GLKD ovaries. R by Pearson’s correlation coefficient. (b) Top, normalized small RNA length profiles: miRNAs (gray columns, insets) and piRNAs/siRNAs (black or red columns) from ovaries with indicated genotypes. Bottom, length distribution and abundance of repeat-derived small RNAs (blue, sense; red, antisense). (c) GFP-Piwi immunostaining from ovaries of indicated genotypes. Scale bars, 10 μm. (d) qPCR result of ova mRNA levels in control and ova GLKD ovaries. Values are means ±SEM.; n = 4. P value by two-tailed Student t-test. (e) Heat map displaying repeat small RNA levels as reads per million (rpm) for individual transposon in ova GLKD and w1118 ovaries. (f) qPCR result of TE mRNA levels in w1118 and tjGAL4 >ova RNAi (shift to 29°C for 7 days) ovaries. ***, p<0.001 by two-tailed Student’s t-test. Values are means ± SEM.; n = 3.