Fig. S6

Figures for Cao et al., 2017
Figure Caption

Fig. S6 Chemical Screening for Regulators of Epicardial Cell Proliferation (Related to Figure 7)
(A) Whole mounted tcf21:nucEGFP ventricles after 6 days of culture as explants, treated with DMSO (Vehicle) or GSK1007102B. Framed regions are enlarged and shown below.
(B) Quantification of EGFP fluorescence density. n = 8 (Vehicle) and 5 (GSK1007102B) explants, respectively. *** P < 0.001, Student’s t-test. Bars indicate mean ± S.D.
(C) Images of fixed and flattened tcf21:nucEGFP explants after 2 days of culture with DMSO (Vehicle) or SB431542. Samples were incubated with EdU for 1 h before fixation (shown in red and grayscale). DNA was stained with DAPI (blue). Scale bars, 100 μm.
(D) Quantification of EGFP+EdU+ nuclei per mm2 on the ventricular surface, from hearts in (C). n = 3 explants for each. * P < 0.05, Student’s t-test. All error bars indicate S.D.
(E, F) PI3K regulates epicardial regeneration. (E) Schematic for experiments in (F). tcf21:NTR; tcf21:nucEGFP animals were used. LY294002 (50 μM) was added for 8 days (3 - 11 dpi). Then, explants were rinsed twice with PBS and further cultured for 4 days. (F) Epicardial regeneration ex vivo in the presence of DMSO (Vehicle) or 50 μm LY294002. All vehicle-treated explants (16 of 16) displayed regeneration. All LY294002- treated explants displayed a block of regeneration (23 of 23), which resumed in each case after drug wash-out. Scale bar, 200 μm.

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Reprinted from Developmental Cell, 42, Cao, J., Wang, J., Jackman, C.P., Cox, A.H., Trembley, M.A., Balowski, J.J., Cox, B.D., De Simone, A., Dickson, A.L., Di Talia, S., Small, E.M., Kiehart, D.P., Bursac, N., Poss, K.D., Tension Creates an Endoreplication Wavefront that Leads Regeneration of Epicardial Tissue, 600-615.e4, Copyright (2017) with permission from Elsevier. Full text @ Dev. Cell