of 10
Developmental Cell, Volume
53
Supplemental Information
Reprogramming Axial Level Identity
to Rescue Neural-Crest-Related
Congenital Heart Defects
Shashank Gandhi, Max Ezin, and Marianne E. Bronner
Supplementary figure titles and legends
Supplementary Figure S1 (related to figure 1, 2): Effect of ablations on
cardiovascular development, and quality control on bulk and single cell RNA
-
seq
data.
(A) (left to right) Primary heart tubes of E3 embryos where the cardiac neural folds
were left in
tact, unilaterally ablated on the left, unilaterally ablated on the right, and
bilaterally ablated. No difference was observed on the effect of left and right unilateral
ablation on primary heart tube looping and elongation. (B) Transverse cross
-
section
th
rough an E6 outflow tract of an embryo where the cardiac neural crest was unilaterally
ablated on the left side exhibits Double Outlet Right Ventricle (DORV). (C
-
D) Hierarchical
clustering (C) and Principal Component Analysis (D) done on the two cardiac an
d three
trunk neural crest libraries prepared for population
-
level transcriptional profiling shows
high correlation between replicates. (E
-
G) Quality control on SMART
-
seq sequencing
data showing the number of genes identified per cell (E, G) and high corre
lation between
mapped and sequencing reads (F). (H) Scatter plot comparing genes enriched in the bulk
and single cell RNA
-
seq datasets. (I) Genes validated in main figure 2 using in situ
hybridization were highly expressed across C1, C2, and C4, with lower
expression levels
in the ectomesenchymal C3.
Supplementary Figure S2 (related to figure 3, 4): Role of Tgif1 in cardiac neural
crest.
(A) Loci for
Tgif1
,
Sox8
, and
Ets1
with gRNA target sites highlighted in red. 2
gRNAs for
Tgif1
, 3 gRNAs for
Sox8
, and 4
gRNAs for
Ets1
were designed. (B
-
C) Loss of
Tgif1 resulted in reduced expression of pan
-
neural crest markers Sox10 (B) and FoxD3
(C), but little difference was observed in the number of cells. (D)
The outflow tract of