Fig. 4

Post-surgery timing of secondary bud development. A Illustration showing surgery to isolate secondary bud (yellow) and anterior region of associated primary bud (orange). Excised tissues in red dashed outline. B, C Post-surgery darkfield images of those bud tissues at day 0 and 6, respectively. Secondary bud is outlined with white dotted line. Portion of primary bud is outlined with yellow dashed line. D Illustration showing surgery performed to isolate a secondary bud alone (yellow). E, F Post-surgery darkfield images of secondary bud tissue at day 0 and 12, respectively. The siphon opened at day 13. G Quantitative results comparing the timing of development to heartbeat and siphon opening during normal budding, secondary/primary bud, and secondary bud alone. dps days post-surgery. Asterisks indicate statistical significance (**P < 0.01). Scale bars = 0.25 mm