ZFIN Image: Wu et al., 2016, Fig. S3

Image

Figure Caption

Fig. S3

MP signaling is activated in endocardial and epicardial cells after injury and transgenic overexpression of bmp2b is sufficient to induce accumulation of nuclear pSmad1/5/8 and expression of the BMP target gene id1. Related to Figure 4.

(A) BMP signaling, as revealed by expression of nuclear pSmad1/5/8, is activated in endocardial cells (marked by fli1a:eGFP expression, arrow) and epicardial cells (marked by ET(krt4:eGFP) expression, arrow) at 3 dpi. Scale bar, 25 µm.

(B) At 21 dpi, hsp70l:nog3; myl7:GFP double transgenic fish heat-shocked using the same regime as shown in Figure 5B display a smaller area of ventricular myocardial tissue (positive for myl7:GFP) than heat-shocked myl7:GFP-only siblings. In contrast, hsp70l:bmp2b; myl7:GFP double transgenics contain more myocardial tissue than myl7:GFP-only controls. Size of the myocardial tissue relative to the whole ventricle normalized to myl7:GFP-only siblings is plotted. Error bars represent SEM. Student’s t test, p = 0.0081 (nog3), p = 0.018 (bmp2b).

(C) Overexpression of bmp2b for 6 days via daily heat-shock is sufficient to induce expression of nuclear pSmad1/5/8 in uninjured hsp70l:bmp2b transgenic hearts but not in heat-shocked wild-type hearts. Scale bar, 50 µm.

(D) RT-PCR reveals that overexpression of bmp2b for 2 days via daily heat-shock is sufficient to enhance the expression of bmp2b and id1, a conserved BMP target gene, in hsp70l:bmp2b transgenic hearts compared to heat-shocked wild-type siblings at 3 dpi. β-actin serves as loading control.

(E) hsp70l:bmp2b transgenic hearts and respective wild-type siblings heat-shocked daily for 6 days do not show different wound sizes at 7 dpi. Error bars represent SEM. Student’s t test, p = 0.9.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image.

Reprinted from Developmental Cell, 36(1), Wu, C.C., Kruse, F., Vasudevarao, M.D., Junker, J.P., Zebrowski, D.C., Fischer, K., Noël, E.S., Grün, D., Berezikov, E., Engel, F.B., van Oudenaarden, A., Weidinger, G., Bakkers, J., Spatially Resolved Genome-wide Transcriptional Profiling Identifies BMP Signaling as Essential Regulator of Zebrafish Cardiomyocyte Regeneration, 36-49, Copyright (2016) with permission from Elsevier. Full text @ Dev. Cell