Figure 4

DNA damage, cell death, and proliferation arrest occur in the pancreas and intestine of wrn^-/- larvae

(A–E) gammaH2AX staining of cryosection. (A) The wrn^+/+ (n = 5) and (B) wrn^-/- (n = 5) larvae at 5 dpf, and (C) wrn^+/+ (n = 6), (D) wrn^-/-_mild (n = 6), and (E) wrn^-/-_severe (n = 6) larvae at 7 dpf.

(F and G) Quantification of gammaH2AX staining signals in pancreas (F) and intestine (G) (corresponding to Figures 4A–4E). The number of samples. wrn^+/+ (n = 5): wrn^-/- (n = 5) at 5 dpf; wrn^+/+ (n = 6): wrn^-/-_mild (n = 6): wrn^-/-_severe (n = 6) at 7 dpf. Statistical significance at 5 dpf was assessed using an unpaired two-tailed Student’s t test, and that at 7 dpf was assessed using one-way ANOVA followed by Tukey’s HSD test. Data are presented as mean ± SD. The detailed mean ± SD are shown in Table S8.

(H–L) EdU labeling of cryosection. (H) The wrn^+/+ (n = 4) and (I) wrn^-/- (n = 6) larvae at 5 dpf, and (J) wrn^+/+ (n = 5), (K) wrn^-/-_mild (n = 6) and (L) wrn^-/-_severe (n = 7) larvae at 7 dpf.

(M and N) Quantification of EdU-labeled signals in pancreas (M) and intestine (N) (corresponding to Figures 4H–4L). The number of samples. wrn^+/+ (n = 4): wrn^-/- (n = 6) at 5 dpf; wrn^+/+ (n = 5): wrn^-/-_mild (n = 6): wrn^-/-_severe (n = 7) at 7 dpf. Statistical significance at 5 dpf was assessed using an unpaired two-tailed Student’s t test, and that at 7 dpf was assessed using one-way ANOVA followed by Tukey’s HSD test. Data are presented as mean ± SD. The detailed mean ± SD are shown in Table S8.

(O–S) TUNEL labeling of cryosection. (O) The wrn^+/+ (n = 5) and (P) wrn^-/- (n = 5) larvae at 5 dpf, and (Q) wrn^+/+ (n = 6), (R) wrn^-/-_mild (n = 6), and (S) wrn^-/-_severe (n = 6) larvae at 7 dpf.

(T and U) Quantification of TUNEL labeled signals in pancreas (T) and intestine (U) (corresponding to Figures 4O–4S). The number of samples. wrn^+/+ (n = 5): wrn^-/- (n = 5) at 5 dpf; wrn^+/+ (n = 6): wrn^-/-_mild (n = 6): wrn^-/-_severe (n = 6) at 7 dpf. Statistical significance at 5 dpf was assessed using the Wilcoxon rank-sum test, and that at 7 dpf was assessed using the Kruskal-Wallis test followed by Dunn’s multiple comparisons test with Holm correction. Statistical analysis for the pancreas at 5 dpf was not performed because both wrn^+/+ and wrn^-/- showed zero values. Data are presented as mean ± SD. The detailed mean ± SD are shown in Table S8.

(A–E, H–L, and O–S) Scale bars, 100 μm. Scale bars in white box, 30 μm.