(A) Release of ENPP-1::mScarlet EVs from the IL2 (inner labial 2) cilia labeled as #1, 2 and 3. The IL2 cilia #1 and #2 are in a state of pre-EV release. The cilium #3 is captured in a post-EV release state with the released ENPP-1::mScarlet EVs forming a “stream” of smaller EVs (small arrowheads) that is preceded by 2 larger EVs located at the front end of the “EV stream” (large arrowheads). See Video S1 for time-lapse imaging of ENPP-1::mScarlet EV protruding from the IL2 cilia. (B) Simultaneous release of PKD-2::GFP EVs (green dashed circles) and ENPP-1::mScarlet EVs (magenta dashed circles) from the CEM (cephalic male) cilia. PKD-2::GFP serves as a marker of the CEM cilia and EVs; Ci – cilia. Images show 3D renderings: on merged images green and magenta colors represent different reporters, on images showing individual channels colors code for Z-depth - 2.5 μm for (A) and 4.0 μm for (B).
Black and white maximum intensity projections for (A-B) and other data related to ENPP-1:mScarlet expression are included in Figure S4. Refer to Video S1 for live imaging of ENPP-1::mScarlet EV release from the IL2 cilia.