Fig. 5: A conserved non-coding element under accelerated evolution in MPI is associated with the downregulation of HIF1A.
From: Experimental evidence for cancer resistance in a bat species
a The phylogeny of seven species examined in this study is used to screen the evolutionarily conserved non-coding elements (CNEs) that are under accelerated evolution in MPI. b CNE143336 is located in the regions with ATAC-seq peaks in MSF and without ATAC-seq peaks in MPI-SF. Rep1 and 2 represent two biological replicates. c Luciferase reporter assay using HEK293T and NIH3T3 cell lines shows that the enhancer activity of CNE143336 is significantly lower in MPI compared to the orthologous sequences in mouse and the Chinese rufous horseshoe bat (RSI). The negative control is the PGL3-promoter empty vector-transfected in HEK293T and NIH3T3. d The regulatory activity of CNE143336 is blocked using CRISPR-dCas9 technology with guide RNAs derived from the sequence of CNE143336. e, f When the regulatory activity of CNE143336 is blocked, the expression level of HIF1A significantly decreases. The numbers of dots (n = 3) in (c), (d), and (f) represent the number of independent experiments. All data are presented as mean ± SD. The P values are from two-tailed Student’s t tests. Source data are provided as a Source Data file.
