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. 2021 Jul 13;10:e62375. doi: 10.7554/eLife.62375

Figure 1. Satellite DNAs (satDNAs) are expressed in ovaries and testes.

(A) Rsp satDNA transcription level in various tissues (corresponding result for 1.688 is shown in Figure 1—figure supplement 1C). Carcass: whole body without the head, reproductive organs, and digestive tract. Source data in Figure 1—source data 1. (B) RNA fluorescence in situ hybridization shows evidence for Rsp and 1.688-derived transcripts in testes and ovaries; asterisk indicates the hub. The probe for 1.688 recognizes all 1.688 subfamilies except for 260-bp on chromosome 2L. (C) Northern blot probed with Rsp. Total RNA was extracted from ovaries of fly lines with varying copy numbers of Rsp: ZW144 (200 copies), Ral357 (600 copies), Iso-1 (1100 copies), Ral380 (2300), and lt pk cn bw (4100). There is no signal after RNaseA treatment. Signal quantification (shown in Figure 1—figure supplement 1D) shows Rsp transcript abundance correlates with its genomic copy number (Pearson’s correlation coefficient r2 = 0.93, p-value=0.02). (D) qPCR and qRT-PCR quantification of Rsp copy number and expression level, respectively, of strains used in northern blot. A linear regression line is shown in the plot with red dotted line (Pearson’s correlation coefficient r2 = 0.98, p-value=0.003). Details for (C) and (D) in Supplementary file 4.

Figure 1—source data 1. Satellite DNAs (SatDNA) transcription level in various developmental stages and tissues.
Related to Figure 1A and Figure 1—figure supplement 1A–C.

Figure 1.

Figure 1—figure supplement 1. Satellite DNAs (satDNAs) are expressed across tissues and developmental stages.

Figure 1—figure supplement 1.

(A, B) SatDNA transcription level in various developmental stages (A for Rsp and B for 1.688). L2: second instar larvae; WPP: white prepupae; Fem: female. Complex satDNA expression increases with adult age. (C) 1.688 satDNA transcription level in various tissues (corresponding result for Rsp is shown in Figure 1A). Carcass: whole body without the head, reproductive organs, and digestive tract. Source data in Figure 1—source data 1. (D) Northern signal quantification of Rsp transcript abundance. Data for (A–C) from modENCODE (Graveley et al., 2011; Brown et al., 2014).
Figure 1—figure supplement 2. RNA fluorescence in situ hybridization (FISH) signals of satellite DNAs (satDNAs) are from RNAs, not DNAs.

Figure 1—figure supplement 2.

RNA FISH of Rsp and 1.688 in ovary (A), with RNaseA treatment prior to probe hybridization (B) and with RNaseH treatment after DNA probe hybridization (C). The 1.688 probe recognizes major 1.688 loci on chromosomes X and 3.