Supplemental Material for Cahoon and Libuda, 2021

Figure S1. Diagram of live imaging mounting and post-imaging recovery. For a comprehensive description see the Methods section “Conditional immobilization for live imaging”.

Table S1. Primer sequences used for CRISPR/Cas9 tagging unc-104, unc-18, and unc-52 with auxin inducible degron tag

Movie
S1. Brightfield 10x timelapse movie of an immobilized hermaphrodite worm. A hermaphrodite worm
containing unc-18::AID*; rgef-1p::TIR1 that was immobilized on 1mM auxin
and mounted under an agar pad. Images were captured every 90 seconds for 60
minutes.

Movie
S2. Autofluorescence in an immobilized L1 worm. A L1 worm containing unc-18::AID*;
rgef-1p::TIR1 that was immobilized on 1mM auxin and mounted under an agar
pad. Timelapse images were taken at 60x magnification and images were captured
every 2 minutes for 60 minutes.

Movie
S3. mCherry::H2B in an immobilized L4 hermaphrodite worm. Timelapse images were
taken at 60x magnification and images were captured every 2 minutes for 44
minutes.

Movie
S4. SYP-2::GFP in an immobilized hermaphrodite worm. Timelapse images were
taken at 60x magnification and images were captured every 5 minutes for 65
minutes. Movie was stabilized for viewing (see Methods for details).

Movie
S5. Whole germline view of SYP-2::GFP in an immobilized hermaphrodite
worm. Timelapse
images were taken at 60x magnification and images were captured every 5 minutes
for 2 hours. The start of the germline occurs out of view of the camera on the
left and germline progresses from left (transition zone/early pachytene) to
right (late pachytene/diplotene) within the movie. Then, the germline makes a U-turn
bend, which is out of view of the camera. The germline comes back into view
indicated by the arrowhead showing the appearance of the diakinesis oocytes on
the bottom right of the movie, which are moving from right to left. Movie was
stabilized for viewing (see Methods for details).

Movie
S6. Brightfield 20x timelapse movie of an immobilized male worm. A male worm containing unc-18::AID*;
rgef-1p::TIR1 that was immobilized on 10mM auxin and mounted under an agar
pad. Images were captured every 90 seconds for 60 minutes.

Movie
S7. SYP-2::GFP in an immobilized male worm. Timelapse images were
taken at 60x magnification and images were captured every 5 minutes for 65
minutes. Movie was stabilized and corrected for photobleaching for viewing (see
Methods for details).

Movie
S8. Whole germline view of SYP-2::GFP in an immobilized male worm. Timelapse images were taken
at 60x magnification and images were captured every 5 minutes for 2 hours. Arrowhead
indicates the approximate start of the germline within the mid-bottom region of
the movie. Germline progression initially starts off moving to the left, then
the germline makes a U-turn bend at the transition zone on the left side of the
movie. After this bend, the germline progression moves from the left (early
pachytene) to right (late pachytene/diplotene) within the movie. The end of the
germline (condensation and mature sperm) occurs farther off to the right
outside of the camera view at this magnification. Movie was stabilized and
corrected for photobleaching for viewing (see Methods for details).