Fig. 2: PATCs promote germline expression from simple extrachromosomal arrays.

a Quantification of germline GFP fluorescence from arrays carrying Ppie-1::fluorescent protein::smu-1 3′-UTR transgenes. The ce-gfp transgenes contain synthetic introns (top) or substitutions with 250 bp and 900 bp endogenous introns with or without PATCs. n = 7, 9, 7, 8, 7, and 9 biologically independent transgenic lines (from top to bottom). b Germline GFP fluorescence from arrays with Psmu-1::gfp transgenes. n = 13, 14, and 5 biologically independent transgenic lines (from top to bottom). c Germline array mCherry fluorescence from a codon-optimized ce-mCherry transgene⁷¹ under control of Psmu-1. n = 11, and 15 biologically independent transgenic lines (from top to bottom). d Germline array fluorescence of Ppie-1::ce-gfp transgenes with single smu-2 introns. n = 11, 10, 10, 10, and 10 biologically independent transgenic lines (from top to bottom). e Germline array fluorescence of Ppie-1::ce-gfp transgenes with intron three from smu-2 at various positions. n = 8, 7, 5, 6, and 4 biologically independent transgenic lines (from top to bottom). f Germline array fluorescence of an N- or C-terminal gfp-tagged smu-1 transgene driven by the Pmex-5 promoter. n = 8 biologically independent transgenic lines for both conditions. g Germline fluorescence as a function of Ppie-1::ce-gfp transgene concentration in simple extrachromosomal arrays (total concentration 100 ng/ul). n = 7 biologically independent transgenic lines for all conditions. All germline fluorescence was quantified from transgenic animals carrying simple extrachromosomal arrays imaged with a ×40 or ×63 oil objective at 25 °C. “gfp” (dark green) refers to the S65C GFP distributed in Fire lab vector kits (A. Fire, unpublished reagents). ce-gfp (light green) refers to a codon-optimized gfp⁴⁴ with piRNA homology removed⁴⁵. PATC-rich transgenes are indicated with a subscript “PATC.” Transgene scale bars = 100 nucleotides. Each datapoint indicates one independent measurement of germline fluorescence scored from 11 animals from an independent transgenic line. Bars indicate the mean, and error bars indicate the SEM. Statistics: a–e Kruskal–Wallis one-way ANOVA. Multiple comparisons: Dunnett’s test. c, f Mann–Whitney two-tailed non-parametric test. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are available in the Source Data file.