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Figure 7 From: Ras-dva small GTPases lost during evolution of amniotes regulate regeneration in anamniotes. Analysis of mitotic cells patterns in amputated tails of tadpoles with normal and inhibited xRas-dva1 or 2 functioning during 1–4 dpa period. (A–C) Transmitted light and fluorescent images of distal areas of 1–4 dpa tails of tadpoles, developed from embryo injected by control MO (A) or xRas-dva2 MO (B) or xRas-dva1 vivoMO (C). (A) Immunostaining with primary rabbit anti-pH3 and secondary anti-rabbit antibodies conjugated with red fluorescent protein CF568 demonstrate gradual increase of mitotic activity in the regenerating area of tadpoles injected by control MO starting from 2 dpa (see B’ for statistics). (B) Transmitted light and fluorescent images of tadpoles injected by xRas-dva2 MO show inhibition of mitotic activity during 2–4 dpa. Scale bar 0,1 mm. (B’) Quantification of pH3-labbeled mitotic cells number in regenerates of 1–4 dpa tadpoles, developed from embryos injected by control, xRas-dva1 or xRas-dva2 MO. Data of five independent experiments (5 tadpoles of each injection type were used in 1 experiment) were used for statistical analysis, statistical significance was determined by paired t test, p < 0,05 (asterisk). Error bars indicate SD. (C) Transmitted light and fluorescent images of tadpoles injected by xRas-dva1 vivoMO show strong inhibition of mitotic activity during 2–3 dpa. (C’) Quantification of mitotic cells number in 1–4 dpa regenerates of tadpoles injected at 0, 1,2 dpa by control, xRas-dva1 or xRas-dva2 vivoMO. Data of five independent experiments (5 tadpoles of each injection type were used in 1 experiment) were used for statistical analysis, statistical significance was determined by paired t test, p < 0,05 (asterisk) Error bars indicate SD. Image published in: Ivanova AS et al. (2018) © The Author(s) 2018. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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