Fig. 3

TOR regulates the early stages of zebrafish heart regeneration

(A) Protein analysis of TOR activity monitored via the phosphorylation of S6 and proliferation via Pcna in regenerating adult zebrafish hearts.

(B) Quantification of pS6/S6 in (A) #p = 0.054, two-tailed t test performed. N = 3 biological replicates with each N being 3–5 pooled hearts.

(C) Protein analysis of TOR activity monitored via the phosphorylation of S6 in regenerating adult zebrafish hearts that have Wnt/β-catenin inhibition through the expression Dkk1b.

(D) Quantification of westerns in (C) ∗∗p < 0.01, two-tailed t test performed. N = 4 biological replicates with each N being 3–5 pooled hearts.

(E) Schematic of adult zebrafish heart ablation and TOR inhibition via rapamycin.

(F) Protein analysis of TOR inhibited regenerating adult zebrafish hearts.

(G) Quantification of protein abundance in (F). ∗∗∗p < 0.001, two-tailed t test performed. N = 3 biological replicates with each N being 3–5 pooled hearts.

(H) Quantification of proliferating cardiomyocytes (Mef2c and Pcna positive cells). One-way ANOVA with pairwise multiple comparison using a Holm-Sidak method were performed. ∗∗p < 0.01. N = 3–5.

(I–K) Immunohistochemistry assessing the number of proliferating cardiomyocytes in the adult zebrafish heart 3 dpi after being treated with no rapamycin and no MTZ, uninjured (I), injured (+MTZ) with no rapamycin (J), or injured (+MTZ) with rapamycin (K). Images have scale bars with a 25 μm length. Blue-DAPI, Red-Pcna, and Green-Mef2c. Yellow arrow heads denote proliferating cardiomyocytes, Pcna and Mef2c positive.

(L) Quantification of TOR positive cardiomyocytes (MF20 and pS6 positive cells). One-way ANOVA with pairwise multiple comparison using a Holm-Sidak method was performed. N = 3, ∗p < 0.05, ∗∗∗p < 0.001. Immunohistochemistry assessing the number of TOR positive cardiomyocytes in the adult zebrafish heart 3 dpi after being treated with no rapamycin and no MTZ, uninjured (M), injured (+MTZ) with no rapamycin (N), or injured (+MTZ) with rapamycin (O). Scale bars: 25 μm. Bar graphs show individual data points with error bars representing standard error. Source data are provided as a Data S2.