Supplemental Material for Camino et al., 2020 Eric Camino Michael Weinstein Mary List Jordan E. Vellky Mark Rebeiz thomas williams 10.25387/g3.10299482.v1 https://gsajournals.figshare.com/articles/figure/Supplemental_Material_for_Camino_et_al_2020/10299482 Supplementary material for Camino et al. manuscript. This includes 2 Supplementary documents and 2 Supplementary figures.<br><b></b><div><p><b>Figure S1. Expression of the DsRed.T4-NLS and EGFP-NLS reporters at the single-cell level. </b>(<b>A</b>) Schematic of the evaluated reporter transgenes. Here, the dimorphic element enhancer is at an equal proximity to the DsRed.T4-NLS (red oval) and EGFP-NLS (green oval) reporter due to the absence of a spacer sequence. Expression of (<b>A’</b>) EGFP-NLS and (<b>A’’</b>) DsRed.T4-NLS is prominent in the female A5 and A6 segments. (<b>A’’’</b>) When the DsRed.T4-NLS and EGFP-NLS expression patterns were merged, widespread co-expression is observed at the single-cell level, indicating that both reporters can be active in the same cells. </p> <p> </p> <b>Figure S2. Promoter competition and spacer sequence regulatory activity had little to no influence on the expressions of the proximal and distal reporter genes. </b>(<b>A</b>) <i>bab</i> locus showing the relative position of the dimorphic element (DE) core enhancer from the promoters (black arrows) for the <i>bab1</i> and <i>bab2</i> genes. (<b>B-E</b>) Schematics of the evaluated DsRed.T4-NLS (red oval) and EGFP-NLS (green oval) reporter transgenes. (<b>B’-E’</b>) Expression of the EGFP-NLS reporter in the abdomens of transgenic female pupae. (<b>B’’-E’’</b>) Expression of the DsRed.T4-NLS reporter in the abdomens of transgenic female pupae. (<b>B</b>) At equal spacing from the reporter genes, (<b>B’</b> and <b>B’’</b>) the dimorphic element drove seemingly identical patterns and comparative levels of EGFP-NLS and DsRed.T4-NLS reporter expression. (<b>C</b>) When separated from the dimorphic element by an 8 kb spacer, (<b>C’</b> and <b>C’’</b>) little to no expression occurred for the distal DsRed.T4-NLS reporter while expression occurred for the proximal EGFP-NLS reporter in the dimorphic element pattern. (<b>D</b>) In the absence of a promoter for the proximal reporter (red X), (<b>D’</b> and <b>D’’</b>) EGFP-NLS expression was absent, and little to no expression occurred from the distal DsRed.T4-NLS reporter. (<b>E</b>) When the dimorphic element enhancer was absent, (<b>E’</b> and <b>E’’</b>) the 8 kb spacer sequence drove little to no expression of the DsRed.T4-NLS and EGFP-NLS reporters. <br></div> 2020-01-02 16:29:27 enhancer promoter Drosophila remote control element tethering element Genetics