A) Illustration of the events that lead to zygote polarization. See text for a
detailed description.
B) Schematic of a SiMPull experiment to analyze the PAR-6/aPKC interaction.
Individual embryos were staged based on morphology, and endogenously tagged
PAR-6::HaloTag was pulled down. The single-molecule images shown are actual data
from pull-downs at the indicated stages. Scale bars represent 5 μm. Note
that the oligomeric complexes (arrowheads) are not macroscopic; they are
diffraction-limited objects, but appear larger because the images were scaled so
that monomers would be visible.
C, D) Measurements of the fraction of PAR-6 molecules in complex with aPKC (C) or
the fraction of aPKC molecules in complex with PAR-6 (D) at the indicated
stages. Each circle in the plots shows the result of one single-cell experiment,
with the size of the circle representing the number of molecules that were
counted in that experiment. The lines show the weighted mean, and error bars
represent 95% confidence intervals.
E) Measurement of the fraction of PAR-6/aPKC heterodimers found in oligomers of
different sizes. The experiment was conducted by pulling down PAR-6::HaloTag and
counting the number of co-precipitated mNG::aPKC molecules in each complex. For
each stage, the distribution of numbers of molecules found in oligomers of
different sizes is shown as a vertical histogram. For clarity, the monomer
fraction (80–90% of total molecules) is not shown. n =
10,138 molecules counted from 7 embryos for pre-polarization phase, n =
5,698 molecules counted from 6 embryos for establishment phase, n =
5,550 molecules counted from 6 embryos for maintenance phase and n =
6,944 molecules counted from 7 embryos for establishment embryos derived from
mothers treated with par-3 RNAi.
F) Images of cortical PAR-6::mNG in live embryos at the indicated stages.
Anterior is to the left. Scale bar represents 10 μm.