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Figure 7. Yap loss of function induces DNA damage.(A) γ-H2AX immunolabeling in the CMZ of retinal sections from stage 40 tadpoles following two-cell stage microinjection of Yap-5-mismatch-MO (control) or Yap-MO. Arrows point to γ-H2AX-positive cells. (B) Corresponding quantification. (C) TUNEL assay on retinal sections from stage 40 tadpoles injected as in (A). Images on the right show higher magnifications of the CMZ delineated with dotted lines. (D) Quantification of TUNEL-positive cells in the different compartments of the CMZ as illustrated on the schematic. (E) 2 days-chase of EdU-labeled cells in the CMZ of stage 42 tadpoles injected as in (A). EdU-positive cells inside the zone encircled with a red dotted line have exited the CMZ (white dotted lines) and integrated the different retinal layers. GCL: ganglion cell layer; INL: inner nuclear layer; PR: photoreceptor layer. (F) Quantification of EdU-positive cells in the neural retina layers. (G) mRNA expression levels of cell cycle genes as measured with the NanoString nCounter system in heads from stage 40 tadpoles following two-cell stage microinjection of Standard MO (control) or Yap-MO. Data are the mean of four independent experiments. (H) In situ hybridization analyses of p53 and p21 expression on retinal sections from stage 40 tadpoles injected as in (A). The number of analyzed retinas per condition is indicated in each bar (B, F) or on the graph (n in D). Data are represented as mean ± SEM. Scale bar = 40 µm.DOI:
http://dx.doi.org/10.7554/eLife.08488.014 |
