XB-IMG-172718
Xenbase Image ID: 172718
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Fig. S5 (related to Fig. 5). Assessment of the activities of Ascl1 on neuronal gene
expression and H3K9me2.
(A) Semi-quantitative PCR analyses of gene expression in animal cap explants treated
with Neurog2 or Ascl1. (B) Western blot detection of overexpressed 6MT-Neurog2
and 6MT-Ascl1 in the animal cap explants prepared through the same procedure as
done in (A). (C, C’) WISH (C) and RT-qPCR (C’) detection of gene expression in
control and Ascl1 splice blocking MO (Ascl1 sMO, 80 ng)-injected embryos at the
stage 18. * P<0.05; ** P<0.01; *** P<0.005, according to two-tailed Student t-test..
(D, E) Anti-H3K9me2 ChIP-qPCR analyses showing that 80 ng Ascl1 sMO did not
alter the H3K9me2 marks on the promoter regions of neurod1 (D) or tubb2b (E). (F)
ChIP-qPCR detection of KDM3A on the -36 bp position of tubb2b promoter.
6MT-neurog2 mRNA (500 pg) and 6MT-ascl1 mRNA (200 pg) were individually
injected at the 2-cell stage and embryos were then harvested at the stage 15 followed
by ChIP-qPCR procedures. (G-I) ChIP-qPCR data showing the effects of ectopic
Neurog2 and Ascl1 on the promoter region of myt1. Both ectopic Neurog2 and Ascl1
were able to bind myt1 promoter (G). Only overexpressed Neurog2 increased the level
of KDM3A (H), and decreased the H3K9me2 marks (I) on the promoter of myt1. *
P<0.05; ** P<0.01; *** P<0.005; ns: no significance, according to two-tailed
Student’s t-test. Image published in: Lin H et al. (2017) Copyright © 2017. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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