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Canonical TGF-β signaling is required for adult zebrafish tendon regeneration. a Schematic of experimental design for inhibition of TGF-β signaling during tendon regeneration. b Representative 2-photon stacks of DMSO and SB-431542-treated regenerating tendons at 7 days post-injury (dpi) in scxa:mCherry zebrafish overlaid with second harmonic generation (SHG) signal and Draq5 (in blue) to label nuclei. The 0-7 dpi treatment and 1-7 dpi treatment are shown in the top and bottom panels, respectively. Yellow arrowheads denote the severed tendon ends. Scale bar, 100 µm. c Quantification of the percentage of scxa:mCherry+ cells detected in the injury site at 7 dpi in DMSO and SB-431542-treated zebrafish for both the 0-7 dpi and 1-7 dpi treatments. Unpaired two-tailed t-tests comparing DMSO controls for each treatment with their respective SB-431542-treated counterparts were performed for statistical analysis (0-7 dpi: DMSO, N = 8; SB, N = 6; 1-7 dpi: DMSO, N = 8; SB, N = 7). ****p < 0.0001. d Quantification of the defect size at 7 dpi in DMSO and SB-431542 (SB)-treated zebrafish in both the 0-7 dpi and 1-7 dpi treatments. Unpaired two-tailed t-tests comparing DMSO controls for each treatment with their respective SB-431542-treated counterparts were performed for statistical analysis (0-7 dpi: DMSO, N = 8; SB, N = 7; 1-7 dpi: DMSO, N = 8; SB, N = 7). ****p < 0.0001. Box and whiskers plots in c and d show whiskers extending from the minimum to maximum values, a line at the median, and a box encompassing the 25th to 75th percentiles.
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