Supplemental Material for Sae-Lee et al., 2020
Wisath Sae-Lee
Luisa L. Scott
lotti brose
Aliyah J. Encarnacion
ted shi
Pragati Kore
Lashaun oyibo
Congxi Ye
Susan K. Rozmiarek
Jonathan T. Pierce
10.25387/g3.12493769.v2
https://gsajournals.figshare.com/articles/dataset/Supplemental_Material_for_Sae-Lee_et_al_2020/12493769
<p></p><p><b>Figure
S1.</b>
Representative images of degenerating HSN neurons. <b>A-C,</b> Degenerated HSN
neurons had dim (arrow heads) or barely visible (dotted circle) cell bodies. <b>D-G,</b>
Earlier evidence of HSN neuron degeneration was visible as blebbing of the cell
body (arrows). <b>H-I</b>, Processes of
degenerating HSN neurons also showed beading (H, arrows) that was not visible
in healthy HSN neurons (I). Autofluorescence from intestine bright in panels H
and I due to higher gain to visualize HSN axon. VC4 and VC5 neurons fluorescing
out of focus in panels B-F. </p>
<p> </p>
<p><b>Figure
S2.</b> APP::mCherry is expressed throughout
the nervous system.<b> A</b> DIC and fluorescent photomicrographs at 40X along
the length of the worm. A C-terminal mCherry tag shows APP expression. As
expected, we found that APP expressed under a pan-neuronal promoter was seen
throughout the entire nervous system including the nerve ring and along the
ventral nerve cord in the absence (left) and presence (right) of exAPOE4. <b>B </b>Enlarged
photomicrographs of the ventral nerve cord taken mid-body show that APP is
expressed in a punctate pattern in the VCs both when APOE4 expression is absent
(top) and present (bottom). <b>C</b> Enlarged photomicrographs show that APP is
expressed in a punctate pattern in the HSNs.</p>
<p> </p>
<p><b>Figure
S3.</b> <i>APOE3
</i>and<i> APOE4</i> transgene
levels. <b><i>A,</i></b> Semi-quantitative analysis of level of <i>APOE</i> transgene arrays in genomic DNA. Average
band intensities of three independent genomic preparations from five
worms/strain following PCR for human APOE presented as mean ±
SEM. No significant differences were found when comparing levels for APOE3 vs
APOE4 strains after correcting for multiple comparisons. <b><i>B,</i></b> Expression
of <i>APOE </i>determined by RT-qPCR of mRNA isolated from 3 (integrated) or 9
(extrachromosomal) near-starved 6-cm plates of worms. DCt
values reveal little difference in gene expression between strains. Values
represent the mean ± SE of two independent
biological replicates with three technical replicates each. Data is represented
as gene expression relative to the reference control gene <i>tba-1</i>. </p>
<p> </p>
<p><b>Table S1</b>. List of strains used
throughout our study arranged by Figure.</p><br><p></p>
2020-07-10 15:11:37
APP
apoe4
C. elegans
neurodegeneration
Neurogenetics