- Title
-
Tnni1b-ECR183-d2, an 87 bp cardiac enhancer of zebrafish
- Authors
- Zhang, Y., Wang, F., Wu, F., Wang, Y., Wang, X., Gui, Y., Li, Q.
- Source
- Full text @ Peer J.
(A) Enhancer activity detection construct used for microinjection. (B) Enhancer activity detection construct used for the luciferase reporter assay. |
(A) ECR Browser and parameter set between zebrafish and human genomes around the 219 kb genome range encompassing the |
Lateral views of zebrafish embryos with heart-specific GFP expression after injection with tol2 mRNA and tnni1b-ECR183 (A–C) or tnni1b-ECR183-d2 (D–F), scale bars = 100 µm. The average ratios of heart-specific GFP expression embryos to the total surviving embryos were 10/45 (22%) at 24 hpf, 15/41 (37%) at 48 hpf, and 15/39 (38%) at 72 hpf in the tnni1b-ECR183 group and 7/59 (12%) at 24 hpf, 18/48 (38%) at 48 hpf and 16/43 (37%) at 72 hpf in the tnni1b-ECR183-d2 group. (G) Rate of embryos with GFP expression in different tissues at 48 hpf after injection with tnni1b-ECR183. (H) Rate of embryos with heart-specific GFP expression from transient injections of tnni1b-ECR183 and each truncated enhancer with tol2 transposase at 48 hpf. A |
TFBSs are displayed in fragments of different colors and numbers. The area division of the triangle represents the distribution density of TFBSs. |
(A–E) Lateral views of zebrafish embryos with heart-specific GFP expression at 24 hpf, 48 hpf, 72 hpf, 96 hpf and 5 dpf. (F) Ventral view of zebrafish embryo with heart-specific GFP expression at 10 dpf. Scale bars = 100 µm. |
(A–F) Lateral views of zebrafish embryos with GFP expression driven by tnni1b-ECR183-d2 (A–C) and those with mCherry expression driven by the promoter of |
Enhancer activity identification of tnni1b-ECR183 and tnni1b-ECR183-d2 by the luciferase assay in the HEK293 (A) and HL1 cell lines (B). (C) Putative TFBS positions and mutations. Analysis of the enhancer activity of tnni1b-ECR183-d2 after putative TFs were overexpressed (D) or after NKX2.5 (E) and JUN (F) binding sites were mutated. One-way ANOVA was used to test for homogeneity of variance, and Bonferroni’s test was used to correct for multiple comparisons (A, B, D). A |
(A–F) Lateral views of zebrafish embryos with GFP expression after injection with tol2 mRNA and tnni1b-ECR183-h179. (G–I) Lateral views of zebrafish embryos with GFP expression after injection with tol2 mRNA and tnni1b-ECR183-h84. The arrowheads indicate GFP expression in the skeletal muscles (A, B, D, F, G, H, I), craniofacial muscles (B, C, D, F) and hearts (E, F), scale bars = 100 µm. Rate of embryos with GFP expression in different tissues after injection with tnni1b-ECR183-h179 (J). Enhancer activity identification of tnni1b-ECR183-h179 and tnni1b-ECR183-h84 in HEK293 cell lines (K). Analysis of the enhancer activity of tnni1b-ECR183-h179 (L) and tnni1b-ECR183-h84 (M) after putative TFs were overexpressed. One-way ANOVA was used to test for homogeneity of variance, and Bonferroni’s test was used to correct for multiple comparisons. * |